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Construction of Lrrc10 Protein Expression Vector

LI Xiao-xia1,2,CHEN Xiang-gui1(1.School of Biotechnology Engineering,Xihua University,Chengdu 610039,China; 2.School of Biotechnology Engineering,Sichuan University of Science & Engineering,Zigong 643000,China)  
To construct pET-Lrrc10 and Express protein of Lrrc10,pMD18-T-Lrrc10 was digested by BamH I and Nco I to get Lrrc10,which was ligated with pET-30a(+) digested by BamH I and Nco I.Then the recombinant was transformed into E.coli strain DH5α.Positive strains were identified by PCR to extract recombinants which were transformed into E.coli strain BL21(DE3) to induce Lrrc10 protein with IPTG at 37℃.The results show that the recombinant plamid was constructed successfully and was named as pET-Lrrc10.After induced by IPTG,the E.coli strain BL21(DE3) transformed by pET-Lrrc10 expressed recombinant protein which was consistent with expected molecular weight.Lrrc10 recombinant protein existed as inclusion body and reached its expression peak after induced by IPTG for four hours.
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