羊种布鲁氏菌Wzt基因的克隆及原核表达

Abstract

Abstract: In order to further study the role of Wzt protein in the synthesis of smooth lipopolysaccharide, the PCR technology was used to amplify the Wzt gene of 759 bp .Then it was ligated into pMD20-T vector.The recombinant plasmid pET-28a-Wzt was transformed into E.coli BL21(DE3) for expression under induction of IPTG. The protein product was analyzed by SDS-PAGE and Western blotting. The results showed that the size of the Wzt gene was 759 bp and the sequence homology was 99.87% compared with the sequence of Brucella melitensis 16 M strain with the access number of AF047478.1 in GenBank,proving that the Wzt gene was successfully cloned. Also, the prokarotic expression vector pET-28a-Wzt was successfully constructed and the Wzt protein with molecular weight of 30 ku was highly expressed in E.coli BL21(DE3), which all the above proved that Wzt gene was successfully expressed.

Key words: Brucella melitensis; Wzt gene; cloning; prokaryotic expression

CLC Number:

PANG Feng, JIA Xiao-xiao, ZHAO Tian-jing, ZHU Hua-pei, XU Kai-lian, GUO Shi-yu, SHI Qiao-yun, RONG Hui, ZHOU Hai-long, WANG Feng-yang. Cloning and Prokaryotic Expression of Wzt Gene of Brucella melitensis[J]. , 2014, 41(11): 54-57.

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Cloning and Prokaryotic Expression of Wzt Gene of Brucella melitensis

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