A. S., Gamal El din
Not Available
Sohair
Not Available
El-Afifi
Not Available
A. S., Sadik
Not Available
Nashwa
Not Available
M.A. Abd El- Mohsen
Not Available
H. M., Abdelmaksoud
Not Available
ABSTRACT
A fragment with a size of 801 bp from the cp gene was amplified from potato virus YN. RNA 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 was through: (1) Insertion cp gene isolated by RT-PCR into PinPointTM Xa-1 vector by ligation and propagated by transformation process in E. coli. (2) Isolation of plasmid DNA, then used restriction enzymes BamH I 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 C° to produce biotinylated protein for E. coli with Mr 22.5 KDa and fusion protein with Mr ~48 KDa consist of biotin tag and CP product of PVYN. (4) The protein was purified by soft linkTM soft release avidin resin. About 2.85 mg/mI of the expressed protein was purified from 1 L of bacterial culture.
Citation
How to cite this article
A. S., Gamal El din, Sohair, El-Afifi, A. S., Sadik, Nashwa, M.A. Abd El- Mohsen and H. M., Abdelmaksoud, 2005. Large Scale Production of Potato Virus Y Necrotic Strain (PVYN) Coat
Protein Through Expression the cp Gene in E. coli. International Journal of Virology, 1: 6-6.
DOI: 10.3923/ijv.2005.6.6
URL: https://scialert.net/abstract/?doi=ijv.2005.6.6
DOI: 10.3923/ijv.2005.6.6
URL: https://scialert.net/abstract/?doi=ijv.2005.6.6