2009•Journal of the South China Agricultural UniversityRequires access
Prokaryotic expression of Banana streak virus ORF I gene and preparation of its antiserum.
He YunWei, Xiu Chen, Ruan XiaoLei, Wenjing Shen, Liu FuXiu, Huaping Li
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Abstract
ORF I gene of Banana streak virus Guangdong(BSV-GD) isolate was amplified from a BSV-GD recombinant plasmid by PCR,and the gene was expressed by being cloned into prokaryote expression vector pET-28b(+).The interesting fusion protein was about 23 000 in size by SDS-PAGE analysis.The product was existed in form of inclusion body.It was dissolved by denaturalization,and the purified protein was obtained by using the histidine labeling kit of N-terminus of the protein.The antiserum was obtained by immunizing healthy rabbits with the purified protein.The results from Western blot and ELISA analysis showed that it's a special antiserum of BSV with high titer which was determined to be 1∶51 200.This study was a base of the research of BSV including ORF I gene's function and the virus detection.