Plum pox potyvirus (PPV), the causal agent of Sharka disease of Prunus inflicts severe crop losses in affected El-Amar area. The virus isolated from EL-Amar apricot trees, was propagated on apricot healthy seedlings. Degenerated oligonucleotide primers were designed to amplify the N-terminal portion of the capsid protein of PPV. The amplified products were cloned into pGEM-T-Easy vector and hybridized to PPV DNA specific probe labeled with Dig-I 1dUTP. DNA sequencing using fluorescent dideoxy nucleotides showed that the capsid protein region of PPV-EA strain had about 65% sequence homology with other strains of PPV and 45% similarity to the CP or PPV-D strain. A PCR fragment coding for the 43 C-terminal amino acids of the Nib and the N-terminal part of the CP (complete variable region plus 19 amino acids of the conserved core) was cloned and expressed into the pQE- 100 plasmid vector. Upon induction, the viral protein coat gene was expressed as 6xHis-tagged PPV/CP fusion protein in E. coli M15 cells. The fusion protein was confirmed by western blot analysis.
Research Article · Volume 1, Issue 1, 4 April 2023 · Open Access
Molecular Cloning and Expression of the Coat Protein Gene of Plum Pox virus El-Amar Strain in E. coli
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Affiliations & Notes ⌄
- 1Microbiology Department, Faculty of Agriculture, Ain Shams University. Shubhra EIkheima, Cairo, Egypt. 2 Plant Virus and Phytoplasma Research Department, Plant Pathology Research Institute, Agricultural Research Center. Giza. Egypt
Article Info ⌄
- Journal:
- Journal of Virological Sciences
- Published:
- 4 April 2023

Abstract
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