A fragment with a size of 801 bp from the CP gene was amplified from Potato virus YN isolated from potato plants cultivated at Monofyia Governorate. RNA was extracted from virus-infected tissues of D. metel leaves by the use of degenerate primers through polymerase chain reaction (PCR). Large-scale amount of PVY (N-Egypt) coat protein produced by gene expression technique in E. coli through: (I) Insertion of CP gene isolated by RT-PCR into Pinpoint Xa-l Vector by ligation and propagation after transformation process in E. coli. (2) Isolation of plasmid DNA. then used restriction enzymes Bam HI and Bgl II to identify clones containing inserts. To confirm the fragment inserted of CP gene sequence. PCR was performed using specific primers for PVYN CP gene. (3) The gene expression was performed using 100 µM IPTG & 2 µM biotin and incubation for 4h / 37 oC to produce biotinylated protein for E. coli with Mr 22.5 KDa and fusion protein with Mr ≈ 48 KDa consists or biotin tag and CP product or PVYN. (4) The protein was purified by softlink™ soft release avidin resin. About 2.85 mg/ml of the expressed protein was purified from I L of bacterial culture.
Research Article · Volume 1, Issue 1, 4 April 2023 · Open Access
Large Scale Production of Potato Cirus Y Necrotic Strain (PVYN) Coat protein through Expression the CP Gene in E. coli
· · · ·
Affiliations & Notes ⌄
- 1Plant Virus and Phytoplasma Research Department. Plant Pathology Research Institute, Agricultural Research Center. Giza-Egypt. 2 Virology Laboratory, Department of Microbiology. Faculty of Agriculture, Ain Shams University, Shubhra El-Kheima, Cairo, Egypt
Article Info ⌄
- Journal:
- Journal of Virological Sciences
- Published:
- 4 April 2023

Abstract
The full text of this article is available as a PDF — use Download PDF above.
Services
Article usage statistics
Loading…
Related articles
- Molecular Variation of Potato Virus Y Isolated from EgyptJournal of Virological Sciences · 2024
- Cloning, Expression and Nucleotide Sequence of Coat Protein Gene of an Egyptian Isolate of Potato Virus y Strain NTN Infecting Potato PlantsJournal of Virological Sciences · 2023
- Biological and Molecular Characterization of Different Isolates of Potato Virus Y (N Group)Journal of Virological Sciences · 2023
- Production of Polyclonal Antibodies against the Recombiant Citrus tristeza virus Coat Protein Expressed in Escherichia coliJournal of Virological Sciences · 2023
- Molecular Cloning and Expression of the Coat Protein Gene of Plum Pox virus El-Amar Strain in E. coliJournal of Virological Sciences · 2023
- Partial characterization of Necrotic ringspot virus on apple in EGYPTJournal of Virological Sciences · 2021
