布鲁氏菌外膜蛋白16基因的克隆及原核表达

Abstract

Abstract: To successfully clone the outer membrane proteins 16 (Omp16) gene and make prokaryotic expression in E.coli, one pair of primers was designed according to B. melitensis M5-90 strain Omp16 gene sequence in GenBank, and then obtained Omp16 gene which was about 507 bp by PCR from the Brucella genome. After purifying, Omp16 gene was inserted into pMD20-T vector to construct recombinant plasmid pMD-Omp16. pMD-Omp16 transformed into E.coli DH5α and identified it by sequencing, then subcloned to vector pET-28a(+). The constructed recombinant plasmid pET-Omp16 was transformed into E.coli BL21(DE3) for expression under induction of IPTG. Lastly, the expression products of recombinant protein His-Omp16 was identified by Western blotting. The results showed that the prokaryotic expression vector was successfully constructed and expressed Omp16 gene in E.coli BL21.

Key words: B.melitensis; Omp16 gene; cloning; prokaryotic expression

CLC Number:

JIA Xiao-xiao, JIAO Han-wei, GUO Shi-yu, SHI Qiao-yun, RONG Hui, ZHANG Jia-ning, ZHU Hua-pei, DU Li, CHENG Ying, WANG Feng-yang. Cloning and Prokaryotic Expression of Outer Membrane Proteins 16 Gene of Brucella melitensis[J]. , 2013, 40(9): 51-54.

References

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Cloning and Prokaryotic Expression of Outer Membrane Proteins 16 Gene of Brucella melitensis

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