In this study,the recombinant cloning vector pGEM2OVP1 was used as template for PCR to get the coding region of VP1 C terminus,and the coding region of PelB signal peptide coding region was added to the upstreamof VP1C,then the fragment was cloned to prokaryotic expression vector to get recombinant expression plasmid pET230a2PelB2VP1C.The VP1Cfragment was replaced by the VP1 coding fragment to get pET230a2PelB2VP1 expression plasmid.The E.Coli BL212(DE3)2plysS was transformed respectively and solublely expressed fused VP1 and VP1 C terminus.Purified VP1 and VP1 C terminus were obtained by NiNT A His Bind Resin affinity chromatography and a single clear band of 3ku and 20 ku appeared respectively in the SDS2PAGE gel.Western blot analysis showed that purified VP1 and VP1 C terminus could react with bovine antiserum against FMDV of serotype O.The consecutive culture of recombinant strains showed pET230aPelB2VP1C and pET230a2PelB2VP1 has excellent stability,which had practical value in soluble protein expression.
GAO Shan-dian
,
CHANG Hui-yun
,
DU Jun-zheng
,
CONG Guo-zheng
,
SHAO Jun-jun
,
LIN Tong
. Construction and Application of an Efficient,Stable Soluble Prokaryotic Expression Vector[J]. Acta Agriculturae Boreali-Sinica, 2009
, 24(6)
: 46
-49
.
DOI: 10.7668/hbnxb.2009.06.009
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