2009•Acta Phytopathologica SinicaRequires access
Prokaryotic expression and preparation of antiserum of Banana streak virus ORF II gene.
He YunWei, Xiu Chen, Ruan XiaoLei, Liu FuXiu, Huaping Li
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Abstract
ORFⅡ gene of Banana streak virus GuangDong isolate(BSV-GD) 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 fusion protein was about 16.5 kDa in size and was soluble with SDS-PAGE analysis.The purified protein was obtained by using the histidine labeling kit of N-terminus of protein.The antiserum was obtained by immunizing healthy rabbits with the purified protein.Western blot and ELISA analysis showed that the special antiserum of BSV possessed high titer,which was tested as 1∶51 200.The study was a base for further research on BSV including ORFⅡ gene function and virus detection.