CLONING AND HIGH-LEVEL EXPRESSION OF Y.PESTIS LCRV GENE IN E.COLI

Aim To study the biology activity of the protective antigen V protein and the pathogenesis of Y.pestis Methods LcrV gene fragment was amplified from Y.pestis strain through PCR.The fragment was identified and cloned into prokaryotic expression vector pBV220 and constructed recombinant plasmid pBV/LcrV.And then temperature regulated inducement expression was used to express LcrV.Results (1)A about 980bp length PCR product was obtain and the sequence was the same as known LcrV gene sequence.(2)The expression product was detected by SDS-PAGE and an expression band about 38×10 3 MW was found.(3) The aim protein representing 38.4% of total cell protein by scanning map analysis (4)Most of the aim protein is in supernatant after ultrasonic crash.Conclusions We obtained LcrV gene and expressed it in Escherichia coli.Most of the expressed product was soluble.