African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12193

Full Length Research Paper

Heterologous expression of βC1 of Chili leaf curl virus in Pichia pastoris

  Saiqa Andleeb*, Imran Amin, Muhammad Shafiq Shahid, Rob W. Briddon and Shahid Mansoor  
National Institute for Biotechnology and Genetic Engineering (NIBGE), P. O. BOX 577, Jhang Road, Faisalabad, Punjab, Pakistan.  
Email: [email protected]

  •  Accepted: 17 June 2010
  •  Published: 22 November 2010

Abstract

 

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 (SDS-PAGE). This is the first study demonstrating the possibility of expression of βC1protein using P. pastoris.

 

Key words: Monopartite begomoviruses, chili leaf curl betasatellite, heterologous expression, Pichia pastoris, betasatellite.

Abbreviation

Abbreviations: SDS-PAGE, Sodium dodecyl sulfate polyacrylamide gel electrophoresis; LB, Luria Bertani; PCR, polymerase chain reaction; IPTG, isopropyl β-D-1-thiogalactopyranoside; YPD, yeast peptone dextrose; MGM, minimal glycerol medium.