2009•Zhongguo shouyi xuebaoRequires access
Prokaryotic expression by low-temperature and establishment of an indirect ELISA assay for the NS1 gene of porcine parvovirus
Zang KeWei, Chunling Zhang, Yong Zou, Chai Jiaqian, Cui YanShun
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Abstract
The completed NS1 gene was amplified from PPV DNA by PCR method.Then the products were cloned into pET32c vector and the sequence was determined.The recombinant plasmid pET32c-NS1was transformed into BL21 competent cells and expressed in super-high level by using low-temperature induction to reduce formation of inclusion body.Western-blot analysis proved the renaturation protein has a good immunoreactivity against PPV antibody.The indirect ELISA method for the detection of PPV specific antibody in procine serum was established,after the optional working circumstances for the ELISA assay(antigen concentration:2.5 mg/L;optimal serum dilution:1∶200) with chessboard titration.The recombinant NS1 can be applied in differential diagnosis of PPV infections.