Temitope Oluwasegun Cephas Faleye1, Johnson Adekunle Adeniji1, 2 *
...EV-C members to generate recombinant VDPVs in the region exist....
Amitha Reena Gomes1, Belamaranahally Muniveerappa Veeregowda2, Sonnahallipura Munivenkatappa Byregowda1, Vinayagamurthy Balamurugan3*
...mals. With the advent of recombinant DNA technology, recombinant protein based vaccines and/or diagnostics are being tested in various heterologous systems across the globe for development of vaccines and/or diagnostic antigens. The recombinant viral proteins, virus like particle based vaccines, bivalent/multivalent vaccines, recombinant viral vectored v...
Wenmin Cheng1,2*, Weirong Pan1,2, Yubo Qing1, Yingchao Liu1, Xingqin Zha1,2, Yan Huang2, Jige Xin1,2, Hongjiang Wei1 and Yangzhi Zeng2
...e (BMI), we obtained the recombinant nucleoplasmin (Npm) by using a prokaryotic expression system and compared the difference on the developmental effects between exogenous Npm and its structural analog, polyglutamic acid (PGA). We chose the pMD18-T vector for Npm expression and transformed E. coli BL21 (DE3) cells to construct a prokaryotic expression system. The results showed that the recombinant Npm protein of 32 kDa cou...
Pam D Luka, Frank N Mwiine, Bitrus Yakubu, Joseph Erume, Ricardo Pérez-Sánchez, Hermann Unger and David Shamaki
...ick antibodies using the recombinant TSGP1 indirect ELISA. Of the samples analysed, 13.4% (442/3,288) showed moderate to high reactivity, suggestive of domestic pigs’ exposure to tick-bite. Antibody reactivity’s were found in eight of the 10 states studied and all the states have favourable climatic conditions for soft ticks’ survival and national parks harbouring wild swine (warthogs) able to maintain the developmental cycle of these ticks. ...
Sheraz Ahmed and Fida Mohammad
...nt program. One hundred recombinant inbred lines (RILs) derived from four F4 populations of Flue Cured Virginia (FCV) tobacco were tested to identify effective selection indices. Experimental material was planted at two locations i.e. Mardan (E-1, E-3 and E-5) and Mansehra (E-2, E-4 and E-6) using alpha lattice design with three replicates during 2012/13, 2013/14 and 2014/15. Heritability in broad sense was generally low for all traits except nicotine and red...
Wesam Hasan Mohamed Mady 1*, Bing Liu2, Dong Huang2, A. Arafa1, M.K. Hassan1, M.M. Aly1, Pucheng Chen2, Yongping Jiang2* and Hualan Chen2
...e in vitro expression of recombinant plasmid DNA was performed by transfection of 293T (HEK) cell line. The HA protein was analyzed using SDS-PAGE followed by Western blot and immunofluorescence assays. The pCAGG-optiH9 vaccine efficacy was evaluated by intramuscular immunization of SPF chickens with different concentrations of plasmid DNA and the sera were collected weekly post vaccination for antibody detection by HI test. All immunized chickens shown high H...
Wesam Hasan Mohamed Mady1*, Bing Liu2, Dong Huang2, Abdel Satar Arafa1, Mohamed Khalifa Hassan1, Mona Mehrez Aly1, Pucheng Chen2, Yongping Jiang2* and Hualan Chen2
...inst antigens encoded by recombinant DNA. The goal of this study was to construct Egy-H5 plasmid DNA-based vaccine against a currently circulating avian influenza virus H5N1 strain in Egypt targeting the HA gene. The HA gene of AIV Egyptian strain H5N1 was codon optimized for chicken biased codon and sub-cloned into pCAGGS mammalian expression vector under the control of chicken β-actin promoter. The construct was transfected into 293T HEK (Human Embryoni...
Furqan Sabir1, Muhammad Tayyab1,*, Bushra Muneer2, Abu Saeed Hashmi1, Ali Raza Awan1, Naeem Rashid3, Muhammad Wasim1 and Sehrish Firyal1
... and characterization of recombinant thermostable phytase from Thermotoga naphthophila (PHYTN). This study was an initial step for the fulfilment of domestic industrial requirement of phytase in Pakistan.The PCR resulted in the amplification of 1.8 kb phytase gene. SDS-PAGE confirmed the size of recombinant protein as 70 kDa. The optimization studies demonstrated the maximal production of
Zhenyu Chang1, Hui Zhang1, Khalid Mehmood1,2, Mujeeb Ur Rehman1, Xiaodan Yuan1, Zhaoyang Li1, Fazul Nabi1, Xiaoxing Wu1, Xinxin Tian1, Xueting Liu1, Jingen Xu3 and Donghai Zhou1,*
...polysaccharide (LPS) and recombinant human (rh) TFF3. Cell proliferation and TFF3 mRNA expression were examined by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assays and real-time quantitative PCR, respectively. The results showed that rhTFF3 effectively prevented the cell injury induced by LPS (P<0.05). A low concentration of LPS (10 µg/mL) down-regulated the expression of TFF3 mRNA, whereas a high concentration of LPS...
Basit Zeshan1,2,*, Mushtaq A. Saleem2,Javed Iqbal Wattoo2, Mohd Mokhtar Arshad1 and Maizan Mohamed1
...truct by sequencing. The recombinant plasmid was identified by restriction enzyme and sequencing analysis. The in vitro expression of the truncated protein was analyzed in E. coli with a molecular weight of 38kDa determined through SDS-PAGE and confirmed by Western blotting. The recombinant truncated protein was then purified from the culture media. The immunogenicity of the protein was studied in an anim...
Sabah Mansoor1, Muhammad Tayyab1,*, Amna Jawad1, Bushra Munir2, Sehrish Firyal1, Ali Raza Awan1, Naeem Rashid3 and Muhammad Wasim1
... and characterization of recombinant α-amylase (AMYSBS) from Geobacillus SBS-4S. AMYSBS exhibited a highest identity of 99.78% with Geobacillus thermoleovorance GTA. E.coli BL21-CodonPlus (DE3) cells were used as host for expression studies of AMYSBS. Recombinant AMYSBS produced as inclusion bodies was transmitted to soluble active form by denaturing the ...
Habib-ur-Rehman1, Masood Ahmed Siddiqui1,*, Abdul Qayyum1, Arifa Bano1 and Naeem Rashid2
Muhammad Shahbaz Aslam1, Iram Gull1, Zaigham Abbas2,* and Muhammad Amin Athar1
...ion on target cells. The recombinant expression of some proteins in E. coli produces inclusion bodies which are misfolded proteins. The objective of this study was soluble expression of IFNα2-Tα1fusion protein in E. coli using pET-SUMO vector and determination of its biological activity. SUMO-IFNα2-Tα1 was successfully expressed in soluble form with IPTG induction to final concentration 0.5 mM at 37oC for 4 h an...
Fangxiao Dong1,2, Xuan Luo1,2, Delu Yuan1,2, Aixin Liang1,2* and Liguo Yang1,2*
...tin DNA vaccine. A novel recombinant plasmid pEGS/2SS-tPA encoding two copies of somatostatin genes fused to tPA signal sequence was succesfully constructed and identified in MCF-7 cells. Subsequently, thirty female mice randomly were divided into three groups, treating with 50 μg pEGS/2SS-tPA (group T1), 50 μg pEGS/2SS (group T2), and 50 μg pEGFP-N1 (Control group) at weeks 0, 2, 4 and 6. Mating with male mice was performed at 2 weeks post fin...
Ghulam Abbas1, Khalid Pervez Akhtar1, Muhammad Ahsan2, Muhammad Jawad Asghar1, Fiaz Ahmad1 and Muhammad Rizwan3*
...imes; mash interspecific recombinants. In this study, 72 mung × mash interspecific recombinant genotypes and a susceptible mungbean variety Mung Kabuli were screened against Yellow Mosaic Disease (YMD) using disease severity ratings by visual scoring of symptoms to calculate percent disease index (PDI). Tested recombinants responded differently to the disease. None of the tested geno...
Yan-Guo Han1, Xiao-Li Peng1, Kai Li1, Yu-He-Tian Zhao1, Xun-Ping Jiang2, Guang-Xin E1, Yong-Ju Zhao1, Jun-Hua Ye3, Li Xu3, Qin-Tao Zhao3 and Yong-Fu Huang1,*
...d the CpG-SS fusion type recombinant plasmid named ptCS/2SS-asd. Twelve ram lambs were randomly divided into three groups, namely, the treatment (Group T1, ptCS/2SS-asd; Group T2, pCpG + ptS/2SS-asd) groups and the control (Group C, pVAX-asd). The vaccine (1 mg/ram) was injected into the rams at weeks 0, 3 and 6 of the study. The rams in Group T1 and T2 showed significantly higher anti-SS antibody, serum growth hormone and IGF-1 concentrations and average body...
Aisha Khalid1, Muhammad Tayyab1,*, Abdual Rauf Shakoori2, Abu Saeed Hashmi1, Tahir Yaqub3, Ali Raza Awan1, Muhammad Wasim1, Sehrish Firyal1, Zaheer Hussain4 and Munir Ahmad5
... and characterization of recombinant thermostable cellulase from Thermotoga naphthophila. PCR using the genomic DNA of T. naphthophila as template resulted in amplification of 1 kb cellulase gene. The amplified cellulase gene was cloned in pTZ57R/T and sub-cloned in pET28a. The expression of recombinant cellulase was analyzed using BL21 CodonPlus (DE3) cells as expression host. The expression studies resulted i...
Muhammad Shahid Nadeem*, Maryam A. Al-Ghamdi and Jalaluddin Azam Khan
...65 was PCR amplified and recombinant plasmid pET22b-Asn was prepared. L-asparaginase was produced in BL21 (DE3) RIL codon plus strain of E. coli as heterologous protein under 0.3mM IPTG. The recombinant enzyme was purified by DEAE-Sephadex colum based anion-exchange chromatography. On SDS-PAGE, the enzyme exhibited a molecular weight of about 36 KDa. Its specific activity was 1650 U per mg of protein with about 5% glu...
Aisha Khalid1, Muhammad Tayyab1,*, Abu Saeed Hashmi1, Tahir Yaqub2, Ali Raza Awan1, Muhammad Wasim1, Shagufta Saeed1, Sehrish Firyal1 and Abdul Rauf Shakoori3
...he maximal production of recombinant thermostable cellulase from Thermotoga naphthophila using BL21-CodonPlus (DE3) cells as expression host and pET28a as expression vector. Effect of various concentration of Isopropyl β-D-1-thiogalactopyranoside (IPTG), post induction time, effect of temperature and pH were examined for the maximal production of recombinant cellulase. The effect of supplementation of LB medium w...
Aziz Ur Rahman* and Syed Mehar Ali Shah
...l comprised 87 rice F5:7 recombinant inbred lines (RILs) along with three check cultivars namely Pakhal, Kashmir Basmati and Fakhr-e-Malakand. The genetic material was planted in alpha lattice design using three replications across different locations namely Peshawar, Swat, Manshera and Charsadda of the Khyber Pakhtunkhwa province of Pakistan during 2017. Pooled analysis of variance indicated significant (p≤0.01) differences among the environments, genotype...
Sheng Niu1, Ali-Raza Jahejo1, Fa-jie Jia1, Xin Li1, Guan-bao Ning1, Ding Zhang1, Hai-li Ma1, Wei-fang Hao2, Wen-wei Gao1, Yu-jun Zhao1, Shi-min Gao1, Jian-hui Li1, Gui-lan Li1, Fang Yan1, Rong-kun Gao1, Huan-chun Chen1,3 and Wen-xia Tian1,*
... with different doses of recombinant glutathione-S-transferase A3 (rGSTA3) proteins. We explored the responses of HDPs in erythrocytes to thiram-induced TD and rGSTA3 protein by quantitative real time PCR (qRT-PCR). The results showed many HDPs expressions were suppressed by thiram-induced TD, and the expressions of HDPs were upregulated by rGSTA3 protein. These findings demonstrated that mRNA expressions of HDPs are highly related to thiram-induced TD, and rG...
Gul Afshan1,2,*, Soumble Zulfiqar2, Sumaira Mehboob2, Muhammad Tahir Javed Khan1 and Abdul Rauf Shakoori2,*
...ed on the development of recombinant vaccine against HCV by utilizing HCV3a envelope glycoprotein E2. HCV3a E2 gene was amplified and cloned first in cloning vector pTG19 and subcloned in expression vector pET21a. E2 protein, expressed in insoluble form, was purified by repeated sonications, followed by denaturation and refolding through fractional dialysis with urea. Multiple alignment with other sequences showed nucleotide variations of HCV3a E2 compa...
Muhammad Ilyas* and Fida Mohammad
...sing 55 F5:8 bread wheat recombinant inbred lines (RILs) with five checks. Experimental material was planted using alpha lattice design in two replicates at Peshawar (E1and E5), Kohat (E2 and E6), Sarai Naurang (E3 and E7) and Dera Ismail Khan (E4 and E8) (Khyber Pakhtunkhwa) during 2014/15 and 2015/16. Locations in each year were considered as independent environments. Pooled ANOVA revealed significant interaction due to GE for days to heading, days to maturi...
Yating Cheng1, Wenlong Shi1, Xue Xiao2, Qirong Zhang2, Qihao Zhang1, Zhijian Su1, Qi Xiang1,2* and Yadong Huang1,2
...structural properties of recombinant collagen by selecting a certain degree of hydroxylation of proline residues, which should contribute to creating engineered collagens with specific end uses.
...
Tahsin Razzaq*, Muhammad Fareed Khan and Shahid Iqbal Awan
...re is improvement in new recombinants of selected parents for yield enhancement. The present study revealed the effectiveness of selection and hybridization for enhancement of resistance to NCLB and improvement of yield and yield associated traits and their utilization in future breeding program.
...
Anam Tariq, Alina Gul, Majida Atta Muhammad, Samia Falak and Naeem Rashid*
...esulted in production of recombinant protein in the cytoplasm which secreted gradually to the extracellular culture medium. Determination of the N-terminal amino acid sequence of the recombinant protein, in the extracellular medium, revealed that the 19 amino acid signal peptide was cleaved between Ala19 and Gly20. It seems probable that the signal peptide of TK0522 can be used for secretion of other
Anand Kushwaha, Amit Kumar, Aparna Madhavan, Durga Goswami, Golmei Poulinlu and Gnanavel Venkatesan*
...lar tools are available, recombinant protein based diagnostic assays namely ELISA is safer and robust to handle large sample size and also to minimize labor/time. However, the genus Capripoxvirus encodes putative 147 proteins in their genome, among which some of them are reported as potential immunogenic candidate genes. Selection and use of such candidate immunogenic proteins from an array of genes located at different structures of a mature virion are the re...
Adewumi Olubusuyi Moses1,2*, Faleye Temitope Oluwasegun Cephas,1,3, Ope-Ewe Oludayo Oluwaseyi1, Ibitoye Ibipeju Henrietta1, Arowosaye Abiola Opeyemi1, Adelowo Oluwatosin Deji1 and Adeniji Johnson Adekunle1,2,4
Ghulam Akbar1*, Muhammad Anjum Zia1, Ali Ahmad2, Neha Arooj1 and Shahneela Nusrat1
Mahmoud Mohamed Ahmed Youssef* and Suzan Abd-Elazeim Hassabo
The Role of Genetic Engineering in Management of Plant Parasitic Nematodes with Emphasis on Root-Knot Nematodes: A Review
...l used as a host for the recombinant DNA technology – based industrial production of proteins and peptides to manage root-knot nematode. Mi gene resistance in tomato plants has been utilized to manage M. incognita and M. javanica. Also, protoplast fusion between Pseudomonas fluorescens and P. aeruginosa to manage M. incognita was utilized. Many genes expressed in nematode feeding cells or the regulatory regions that control these genes have been isolated...
Saba Aleem1*, Mehvish Tahir2, Iram Sharif3, Muqadas Aleem4, Muhammad Najeebullah2, Ali Nawaz1, Amina Batool1, Muhammad Imran Khan1 and Waheed Arshad1
Principal Component and Cluster Analyses as Tools in the Assessment of Genetic Diversity for Late Season Cauliflower Genotypes
...ed for obtaining diverse recombinants in segregating generations, exploiting heterosis, and broaden the genetic base of the cauliflower germplasm....
Hesham Saeed1*, Manal Abdel-Fattah1, Ahmad Eldoksh1, Farid S. Ataya2 and Manal Shalaby3
...n with IPTG for 5 h. The recombinant IFNα was expressed in the form of inclusion bodies that were separated and solubilized in vitro and the protein was refolded using SDS and KCl. The folded protein is then purified using on Ni-NTA Agarose affinity chromatography and the purity was judged by SDS-PAGE. Moreover, the effect of the recombinant IFNα of the viability of cancer cell line was assessed by MTT ass...
RehamM. El-Bakrey1, Shimaa M.G. Mansour2, Mohammed A. El Sisi1 AndAmal A.M. Eid1
...11. Both inactivated and recombinant fowlpox virus expressing H5 (rFP-AI-H5) vaccines were measured. Vaccine efficacy was evaluated by percentage of protection from mortality, morbidity and reduction in virus shedding from respiratory and/orintestinal tracts. Despite the H5 antibody responses in vaccinated chickens with program I (rFP-AI-H5" vaccine then Re-5) being significantly lower prior to challenge, it provided good protection (73.3%) against the le...
Aya El-Turkey1, A. K. El-Attar1, A. E. Aboulata1, B. Othman2 and K. A. El-dougdoug2
...s liberated from 1 μg recombinant plasmid by using the restriction endonuclease EcoRI. HSV-2gD was successfully sub-cloned into binary plant expression vector PBI-121. HSV-2gD subunit-containing binary vector PBI 121 were transformed into Agrobacterium tumifaciens LBA 4404 strain for Agro-inoculation. Tomato plants were transferred into regeneration medium and genetically transformed with the HSV-2gD-PB21 binary vector through Agrobacterium co cultivation. ...
A. A. Rezk1,3, K. A. Alhudaib2 and A. M. Soliman2,3
Ola Youssef 1; EL-Deeb A.H.2 Nassif S.A.1 and Ahmed A. El-Sanousi 2.
...otocol for evaluation of recombinant H5
vaccines had been established yet so, these vaccines are evaluated by applying the conventional QC
tests such as identity, safety, purity, potency and efficacy that were not satisfactory in vaccine
evaluation.
Objective: The present study was conducted to standardize a protocol for quality control and
evaluation of the recombinant...
Aly M. Abdel-Salam
...ould be produced through recombinant protein system for successful evaluation of resistance....
Lamiaa M. Omar, Mohamed A. Abdrabo, Dali M. Omar, Nermeen A. Marden
...
inactivated recombinant vaccines respectively. While, the ranges from 102, 102 and 10 when measured
by rRT-PCR respectively. Also, the live recombinant vaccine could not reduce the viral shedding
when measured by both egg inoculation and rRT-PCR.
Conclusion: Depending on the previous phenomena this study showed that despite the expensive and
latency of VI, it is mo...
Bingjie Zhou1, Hitesh Bhagavanbhai Mangukiya1, Siva Bharath Merugu1, Fakhar-un-Nisa Yunus1, Yuchen Fan1, Zhenghua Wu1,* and Dawei Li1,2,*
...auto-fluorescent his-tag recombinant AGR2-DsRed (hAD) protein was constructed by linking human AGR2 C-terminus with the red fluorescent protein derived from Discosoma sp. (DsRed) througha flexible linker. It was expressed successfully in Escherichia coli (E. coli) with optimized strain and cultivation parameters. This protein was expressed and purified by Nickel-nitroacetic acid (Ni-NTA) affinity chromatography. The bioactivity of AGR2 was...
Amjed Ali1, Muhammad Tayyab1,*, Sehrish Firyal1, Abu Saeed Hashmi2, Asif Nadeem3, Shagufta Saeed1, Ali Raza Awan1 and Muhammad Wasim1
...gher level production of recombinant β-lactamase from Bacillus subtilis R5. Supplementation of LB medium with various carbon sources including wheat bran, rice bran and molasses could increase the enzyme production from 25 to 43.7, 44.3 and 49 µmole/min whereas the nitrogen sources including tryptone, peptone and yeast extract could enhance the enzyme production from 25 to 40.21, 41 and 43.76 µmole/min, respectively. The highest β-...
Sadaf Ashraf1, Masood Ahmed Siddiqui1*, Kanwal Nisa1, Samar Ali1 and Naeem Rashid2
Lin Huang1, Ling Mai2, Keyan Zhong1 and Xinjun Chen3*
... (SeFABP) and obtain its recombinant protein, the basic physio-biochemistry characteristics, signal peptides, antigen epitopes, transmembrane domains, secondary and tertiary structures, multi sequence alignment and molecular evolutionary tree of SeFABP were predicted and analyzed. On this basis, SeFABP was whole-genome synthesized and cloned into prokaryotic expression vector. The recombinant plasmid pET-30a(+)-SeFABP was co...
Sana Shakoor, Tahir Rehman Samiullah, Naila Shahid, Abdul Qayyum Rao*, Aneela Yasmeen, Sana Tahir, Ayesha Latif, Saira Azam, Ahmad Ali Shahid and Tayyab Husnain
Anis Suraya Mohamad Abir1, Sakinah Yusof1, Abd Rahaman Yasmin1,2, Abdul Rahman Omar1,3, Kok Lian Ho4, Wen Siang Tan1,5, Abdul Razak Mariatulqabtiah1,6*
...BM-DCs) and FWPV and its recombinant viruses are limited compared to other avian viral infectious diseases. Therefore, this study characterised the response of chBM-DCs to infection with the wild-type (WT) FP9 strain of FWPV compared to a recombinant FWPV carrying the H5 gene of avian influenza virus (rFWPV-H5) using morphological and immunophenotypical observations. The results showed that the rFWPV-H5-infected chBM-DCs for...
Amjed Ali1, Muhammad Tayyab1*, Abu Saeed Hashmi2, Asif Nadeem3, Shumaila Hanif4, Sehrish Firyal1, Shagufta Saeed1, Ali Raza Awan1 and Muhammad Wasim1
... the characterization of recombinant β-lactamase from a locally isolated strain Bacillus subtilis R5. The study was an initial step for the fulfillment of commercial needs of β-lactamase in Pakistan. The 1 kb β-lactamase gene was amplified by PCR using the genomic DNA of B. subtilis R5 as template. The purified PCR product was cloned in pTZ57R/T and sub-cloned in pET21a. Expression of recombinant protein was e...
Amreen Zahra1*, Mushtaq A. Saleem2, Hasnain Javed3, Muhammad Azmat Ullah Khan4 and Abdul Rauf Shakoori4
...G, Subtype C a potential recombinant had 3.33% and unique recombinant form URFs accounted for 13.3% in the HIV Gag sequences. All Pol (URFs) demonstrated 40% of ratio among the isolates, and Subtype D with 6.66%. The present study shows that multiple subtypes of HIV-1 are present in the affected population. Hence, this study highlights the demand of continuous future studies on disease surveillance coupled with responsible e...
Liyun Chang1, Aiju Liu2, Jianshuang Zhang3, Yingbin Chen1 and Zhiyong Liu4*
...and induced to express a recombinant protein (SAG2), identified using SDS-PAGE and Western blot analysis. The purified protein was then used as a coating antigen to establish an indirect ELISA method for detecting the T. gondii antibody in pet cats, whose reaction conditions were optimized. The SAG2 gene was successfully cloned into a pET-21a (+) prokaryotic expression vector, and the recombinant plasmid pET-21a-SAG2 was obt...
Zahra Naz1,2, Fouzia Ismat1, Muhammad Saleem2, Mazhar Iqbal1, Aamir Shehzad1 and Moazur Rahman1,2*
...ted here reveal that the recombinant detergent-solubilized M protein is an active, promising diagnostic antigen and can be exploited in an indirect ELISA for rapid and reliable detection of anti-NDV antibodies in chickens. Owing to the conserved nature of the M protein and the ease of extraction and solubilization in a single step using LDAO, the developed ELISA can be up-scaled for rapid and reliable detection of anti-NDV antibodies and/or NDV infection in a ...
Anara Ryskeldina, Indira Iskakova, Nurgul Sarina , Alexander Shevtsov , Laura Syzdykova, Alexander Shustov, Yerlan Ramankulov, Marat Kuibagarov*
...is work was to produce a recombinant boPAG1 antigen and obtain monoclonal antibodies (mAbs) against boPAG1. We have obtained the boPAG1 cDNA and are reporting its nucleotide sequence. Bacterial expression of a portion of the natural gene encoding a mature form of boPAG1 failed. But a fusion protein made up of E. coli thioredoxin and boPAG1 was efficiently expressed in E. coli. Using the recombinant protein as an antigen, mAb...
Iram Amin1,2*, Shazia Rafique1, Nadeem Ahmed1, Muhammad Shahid1, Samia Afzal1, Tahir Rehman Samiullah1, Mohsin Ahmed Khan1 and Muhammad Idrees1,3
...he study. This antigenic recombinant HDAg protein can be useful for both vaccine development and as a diagnostic marker of HDV. After determination of HDAg antigenic region of HDV and its amplification, the fragment was cloned in bacterial expression vector. The expression of recombinant protein was checked by SDS-PAGE and after the purification of protein by using Ni-affinity column. antigenic protein was confirmed with Wes...
Hagar Magdy Ahmed1, Mohamed Mahrous Amer2*, Khaled Mohamed Elbayoumi1 , Sameh Abdel- Moez Ahmed Amer1, Asmaa Mahmoud Matoaq1 , Mohamed Abdel Aziz Kutkat1, Gomaa Abd El-Rhim Abdel-Alim2
...s (genotype II) and live recombinant herpes virus of turkey (rHVT-ND-IBD) alone or in combination with inactivated NDV vaccines either of genotypes II or VII (commercially available or autogenously prepared). Different vaccination regimes are applied at various designated days to 260 commercial broiler chickens in 13 groups; 20 birds each. At 28 days-old, all groups were challenged with NDV genotype VII 1.1 strain “NDV-CHICKEN-EGY-ALEX-NRC-2020” to...
Marat Kuibagarov1, Assylbek Zhylkibayev1, Dinara Kamalova1, Anara Ryskeldina1, Nurdina Yerzhanova1, Yerlan Ramankulov1, Alexandr Shevtsov1*, John A Angelos2
... to prevent IBK based on recombinant pathogenic factors have been evaluated as alternatives to whole cell vaccines. Pilin is one candidate antigen for M. bovoculi, and is a key pathogenic factor necessary for attachment of M. bovis to the surface of the eye epithelium. To evaluate pilA diversity amongst Kazakh isolates of M. bovoculi, the pilA gene sequence was determined for 35 isolates of M. bovoculi. Ten genotypes (gNA1-gNA10) were identified which were 96...
Monu Karki, Amit Kumar and Gnanavel Venkatesan*
...rals, viral vectors, and recombinant vaccines....
Muhammad Shahid Nadeem*, Jalaluddin Azam Khan and Firoz Anwar
...study we have produced a recombinant of ALT from Pyrococcus abyssi in BL21 (DE3) strain of E. coli. The recombinant enzyme was purified by anion exchange chromatography, it displayed a 45kDa band on SDS-PAGE, with 58.1% final recovery, 15.3 fold purification and 139 U/mg specific activity. Maximum enzyme activity was found at pH 8 and above 90C, its KM and Vmax values were found 25µM L-alanine and 149 U/min/mg respecti...
Aadil Sultan, Mohsin Ahmad Khan*, Nadeem Ahmed, Muhammad Hassan, Rashid Bhatti, Hafsa Naeem, Muhammad Islam Khan, Saad Tahir, Samia Afzal* and Ahmad Ali Shahid
...to enhance production of recombinant proteins in bacterial expression system....
Atena Azarniush1, Majid Moghbeli2*, Farshid Kafilzadeh1, Mohammad Kargar1 and Hooshang Jamali1
...ative treatment, and the recombinant Interleukin 11 (IL-11). In this study, IL-11 gene owning two restriction enzyme sites of BamHI and XbaI with a final size of 609 bp was synthesized by GenRay (China) and delivered to pGS vector. The vector was transferred to DH5α, then the gene was subcloned onto pHT43 shuttle vector and transformed into the Bacillus subtilis WB600. The vector containing the cloned gene was induced within the WB600 by IPTG. The SDS-PA...
Beenish Shahid1* and Muhammad Ijaz Khan2
...oestradiol (2 μg/ml), recombinant human follicle stimulating hormone (0.05 IU/ml), human chorionic gonadotrophin (2 IU/ml) and insulin (0.12 IU/ml) alone or together in different combinations at intervals of 8, 16 and 24 h of incubation period during in vitro maturation on buffalo oocyte. The supplementation of TCM-199 with E2+rhFSH showed a highly significant increase in the diameter and maturation of oocytes (P<0.0001), as a greater number of the oocyt...
Burarah Arooj1, Zeeshan Mutahir1, Moazzam Ali1, Mohsina Akhter2, Malik Siddique Mahmood1, Arslan Hamid3 and Mahjabeen Saleem1*
... were used to purify the recombinant 52 kDa protein. Purified chitinase showed optimum activity at 60°C and pH 5.5 with colloidal chitin breakdown. Enzyme showed stable residual activity within pH range of 4.5–6.5 and >70% thermal stability upto 60°C for 2.5 h. The activity of chitinase increased in the presence of Mn+2, SDS, and methanol. Chitinase has Km and Vmax values of 2.287 mg/ml and 6.784 µM/min, respectively, towards colloidal c...
Nguyen Thi Thu Hien1, Dong Huu Rin2, Nguyen Xuan Hoa1, Le Viet Tuan Khanh2, Phung Thang Long1*, Dinh Thi Bich Lan1
...eg;) strong detected the recombinant M. hyopneumoniae P36 protein by ELISA, demonstrating that expressed recombinant P36 protein has the antigenicity and is specific for M. hyopneumoniae. In summary, we have successfully cloned the gene (432 bp) encoding the M. hyopneumoniae P36 protein. The expressed recombinant M. hyopneumoniae P36 protein in strain E. coli BL21 has the antigenicity and ...
Abida Mushtaque1, Ali Ahmad Sheikh1, Aamir Ghafoor1*, Wasim Shehzad2 and Nadeem Ahmad3
... mice. The expression of recombinant OmpH protein (rOmpH) resulted in its accumulation in cytosol. The purified rOmpH protein had a molecular weight of 36 kDa and concentration around 0.973 mg/mL. The BALB/c mice group injected with 100 ng of purified rOmpH exhibited significant protection (80%) when challenged with LD50 of P. multocida B:2. From the obtained results it is evident that rOmpH can be used as subunit vaccine against P. multocida B:2 and the purif...
Soliman*, Ahmed M.; Mahmoud**, Sabry Y. M. and Dawood*, Rehab A.
.... coli) strain DH5a. The recombinant plasmids were obtained and sequenced. The nucleotide sequences were compared with corresponding viral nucleotide sequences reported in GenBank. The sequence analysis showed that; nucleotide sequence of OYDV-EG [Egyptian isolate (HM473189)] had 82-96% similarity compared with ten reported OYDV isolates. Thus, a method of identification and detection by RT-PCR of OYDV was established. This study also aimed to obtain OYDV-free...
*El-Helaly, Sahar H.; ** Ahmed, Amal A.; * Awad, M.A. and ** Soliman, A.M.'
.... coli) strain DH5a. The recombinant plasmids were obtained and sequenced. The nucleotide sequences were compared with corresponding viral nucleotide sequences reported in GenBank. The analysis showed that nucleotide sequence of the Egyptian isolate of AMV (GenBank Accession No. HQ288892) had percentage of similarity ranged from 97-98% with eight reported AMV isolates. Thus, a method of identification and detection by RT-PCR of AMV was established....
El-Sabagh, I.M.; Hussein, H.A.; Amer, H.M.; El-Sanousi, A.A.; Reda,I.M. and M.A. Shalaby
...e polyhedrin promotor. A recombinant baculovirus carrying the VP7 gene was constucted through homologous recombination between the baculovirus transfer vector carrying the VP7 gene and Autographa californica Nuclear Polyhedrosis Virus (AcNPV). Infection of Spodoptera frugiperda (SD) cells with Baculovirus recombinants expressing VP7 protein revealed high reactivity with hyperimmune antiserum to BRV when tested by immunoflure...
Amer, H.M.; Hussein, H. A.; El-Sabagh, I. M.; El-Sanousi, A. A. Saber,M.S. and Shalaby, M. A.
...colonies that harbor the recombinant plasmids carrying N gene in correct orientation. Generation of recombinant baculoviruses was achieved by co-transfection of Spodopetra frugiperda (Sf-9) insect cells with a linearized replication-defective baculovirus DNA (Bac-N-BlueTM) and the transfer vector. The recombinant baculoviruses were plaque purified; verified for the presence of target seque...
Nurlan Akhmetsadykov1*, Tanatar Kydyrov2, Moldir Akhmetzhanova1, Gulnazi Akhmetova3, Maxat Berdikulov4
...s study was to produce a recombinant GM6 antigen derived from T. evansi and investigate its immunological properties for the detection and diagnosis of trypanosomosis. To create a recombinant antigen, bioinformatic analysis of the primary structure of T. evansi GM6 antigen from several regions was carried out. The protein sequence was reverse translated to the nucleotide sequence, after which the gene was synthesized using s...
Siqiang Li1,2, Tiantian Wang1, Peng Sun1, Airong Gao1, Xin Gong1, Yuanhong Xu2, Baogen Wang2, Jun Wu1* and Bo Liu1*
...strain GS115 harboring a recombinant MDS I derived from Trichoderma reesei was constructed via conventional molecular cloning methods, and expressed in a 5-liter fermentation tank. The target protein was purified in three-step purification and identified by peptide mass of fingerprint. The enzymatic activity and optimal reaction conditions of MDS I were detected using DNA sequencer-assisted fluorophore-assisted carbohydrate electrophoresis. We obtained MDS I w...
Muhammad Nadeem1, Muhammad Naveed2,3*, Muhammad Shafiq2, Irfan Rasool2 and Muhammad Afzal Zahid2
...yield and wilt resistant recombinants in subsequent generations. Resultantly, a uniform and high yielding progeny line PCK-09012, later designated as “Noor-2019” was selected and evaluated for yield, agronomy, and insect pests. This cultivar revealed a potential yield of 2882 kg ha-1 in testing phase and outperformed standard checks by producing 78.2%, 13.1%, and 28.8% higher grain yield in station, adaptation and national uniform yield trials, res...
Raza Ali Mangi1,2, Ali Raza Jahejo1,2, Afrasyab Khan1,2, Meng-li Qiao1,2, Muhammad Farhan Qadir1,2, Mazhar Hussain Mangi3, Shi-xiong Yang1,2, Xin-yu Han1,2, Sheng Niu1,2, Ding Zhang1,2, Ying Wang1,2 and Wen-xia Tian1,2*
...o evaluate the effect of recombinant glutathione-S-transferase A3 (rGSTA3) protein on HDR and PRRs in the thiram-induced TD chicken. One hundred twenty arbor acres (AA+) broiler chickens of seven-day-old were equally divided into six groups. Group A, B, and C were treated with 0, 20, 50 μg.kg-1 of rGSTA3 protein, respectively. Group D, E and F were treated with 0, 20, 50 μg.kg-1 of rGSTA3 protein + thiram 100 μg.kg-1. The results showed that chicks tr...
Kashif Abdaal, Aneeza Batool, Muhammad Tariq Navid*, Saeed Ahmed, Asma Saleem Qazi, Waseem Safdar, Haidar Ali, Mobeen Ur Rashid, Somia Rafaqat
...gies including mRNA- and recombinant DNA technologies along with viral vector-based vaccines containing specific DNA segment(s) of an infectious pathogen(s), nanoparticle-based and plant-based delivery mechanisms have proven as more targeted and adaptable approaches to boost immune responses. Hence, it proves the vaccines as a big breakthrough in preventive and therapeutic medicine over 200 years and named “Future Medicine”. To assess vaccine safet...
Muhammad Wasif Gulzar*, Riffat Maqsood, Muhammad Zain, Muhammad Suleman, Tayyab Ur Rehman, Sana Asif, Abdul Wadood and Jawad Hussain
..., vaccinations with DNA, recombinant vaccines, and live, attenuated, or inactivated viruses can be administered. This study provides comprehensive information on immunization, treatment techniques, pathophysiology, epidemiology, transmission, and relevant preventive and control measures....
Quratulain Amjad1,2 and Abdul Rauf Shakoori1,2*
Niaz Hussain1, Muneer Abbas1*, Abdul Ghaffar1, Muhammad Aslam1, Mudassar Khaliq1, Khalid Hussain2, Muahammad Nadeem1, Muhammad Irshad1, Zubeda Parveen1, Fiaz Hussain3 and Azhar Mahmood Aulakh4
...thod, mainly focusing on recombinants possessing high yield potential with bold grains. Uniform lines were selected from F6 generation and bulked. Ultimately, the promising and uniform line “TG1410” later on named “Thal-2020” was selected from F7 population during 2013-14. This selected line underwent rigorous evaluation for yield potential and disease resistance during the process of evolution. In various station, adaptation, and natio...
Aisha Khalid1,2, Roheela Yasmeen1*, Farah Ahmad1, Mudassar Aslam2, Muhammad Tayyab2
...tigated the effects of a recombinant thermostable cellulase from Thermotoga naphthophila (CELTN) on the growth performance of broiler chicks when added to corn-based feed. The thermostable cellulase was produced using BL21 CodonPlus cells with the pET28a-Cellu vector under optimized conditions. A total of 150 one-day-old, unsexed broiler chicks were divided into five treatment groups, each further subdivided into three replicates of 10 chicks with an initial a...
Ningyu Zhu, Runzhen He, Qianrong Liang, Xiaoye Zheng, Gaohua Yao, Wenjun Ma and Xueyan Ding*
...otein gene of MSRV and a recombinant plasmid containing the target gene was constructed as a standard control. The correlation coefficient of the standard curve was 0.998, which indicated a good linear relationship between initial templates and Ct values. The established SYBR Green I RT-qPCR assay had a detection limit of 2.78×101 copies/μL, which was 100 times more sensitive than the conventional PCR. Moreover, the coefficient of variations was less ...
Rakhtasha Munir1, Shazia Rafique2*, Iram Amin1,2, Sameen Ahmed2, Ayesha Vajeeha2, Amjad Ali3, Muhammad Shahid2, Muhammad Umer Khan4 and Muhammad Idrees2
Gamal Abdelaziz 1,2*; Motaleb, M.A.1; Farouk, N.1; Adli A. Selim1*
Novel Research in Microbiology Journal (2019), 3(3): 341-350
...for tumor targeting. New recombinant insulin analogue was successfully produced from E. coli by recombinant DNA technique, and was well labeled with Technetium-99m with a high radiochemical yield of 93.3 ± 2.1 %. Moreover, it showed good in-vitro stability in both saline and human serum. Preclinical evaluation of Technetium-99m [99mTc] Tc-insulin in solid tumor-bearing mice showed high accumulation in tumor tissues. T...
Gamal, Abdelaziz1*; Heba, A. Mostafa2
Novel Research in Microbiology Journal (2020), 4(3): 736-745
...ated virus, subunits and recombinant proteins. However, scientists believe that under the best condition, the vaccine will not be available before January, 2021. The aims of this review were to express the main trials for COVID-19 treatment via drug repositioning, as well as the movement of the companies and the different organizations towards finding a suitable active vaccine, before the crisis become improbable....
Hayam Albalawi1,2, Muhammad Shahid Nadeem1*, Hisham N. Altayeb1,
Saima Iftikhar3, Mariam A. Al-Ghamdi1,4,5, Jalaluddin Azam Khan1 and Ahmed Osman1
...e II. We have produced a recombinant of α-glucosidase from Thermococcus radiotolerans in E. coli strain BL21 (DE3) codon plus RIPL in the presence of 1 mM IPTG. The enzyme was purified by selective precipitation and cation exchange chromatography, it displayed a 30 kDa band on SDS-PAGE. The purified recombinant of enzyme exhibited an optimal activity at pH 6, and 85°C, it retained more than 50% activity after incub...
