Heterologous expression of βC1 of chili leaf curl virus in pichia pastoris

Saiqa Andleeb*, Imran Amin, Muhammad Shafiq Shahid, Rob W. Briddon, Shahid Mansoor

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)

Abstract

Chili pepper is naturally susceptible to a wide range of viruses in all south Asian countries including Pakistan. Chili leaf curl virus is a monopartite begomovirus having single stranded circular betasatellite. It has one open reading frame βC1, required for pathogenicity determined, symptom induction and viral accumulation. It produces viral symptoms like mosaic, mottling, leaf distortion, vein etching, yellowing, stunting and narrowing of leaves. This study was conducted on the basis of βC1 protein, whether it was expressed in prokaryotic and yeast expression system or not because many viral proteins are lethal for the host organism. For this study, specific set of primers for βC1 were designed and amplified product was inserted into pET32a(+) bacterial and pPIC3.5K Pichia vectors for its expression. βC1 was not expressed in BL21 Escherichia coli expression system, while it was expressed in Pichia pastoris, when it was integrated into the genome through electroporation, and expressed protein was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDSPAGE). This is the first study demonstrating the possibility of expression of βC1protein using P. pastoris.

Original languageEnglish
Pages (from-to)8023-8031
Number of pages9
JournalAfrican Journal of Biotechnology
Volume9
Issue number47
DOIs
Publication statusPublished - Nov 22 2010
Externally publishedYes

Keywords

  • Betasatellite
  • Chili leaf curl betasatellite
  • Heterologous expression
  • Monopartite begomoviruses
  • Pichia pastoris

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Genetics
  • Agronomy and Crop Science

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