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Cloning Expression and Characterization of a Bi-functional Antibacterial Peptides CM4 and hsBAFF Fusion Protein in Escherichia Coli(PDF)

《南京师大学报（自然科学版）》[ISSN:1001-4616/CN:32-1239/N]

Issue:
2008年02期
Page:
87-91
Research Field:
生命科学
Publishing date:

Info

Title:
Cloning Expression and Characterization of a Bi-functional Antibacterial Peptides CM4 and hsBAFF Fusion Protein in Escherichia Coli
Author(s):
Liu Haifeng; Liu Ping; Li Nannan; Zhang Shuangquan
Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology,School of Life Science,Nanjing Normal University,Nanjing 210097,China
Keywords:
Antibacter ia l peptide CM4;  hsBAFF;  fusion gene;  E. co li;  expression
PACS:
R346
DOI:
-
Abstract:
The cDNA o f hum an so lub le B lymphocyte stim ulato r ( hsBAFF) w as am plified by PCR from pET30a ( + ) and the gene o f an tibac terial peptide CM 4 by rPCR. The fusion gene o f hsBAFF and CM4 w as am plified by us ing over- lap PCR. The prokaryotic expression plasm id pET30a ( + ) /hsBAFF - CM 4 w as constructed w ith recomb inant DNA techniques after pur ify ing and iden tify ing the DNA fragm ent. Then the plasm id pET30a( + ) /hsBAFF- CM4 w as trans form ed in to BL21( DE3) cells and the expression w as optim ized w ith proper induc ing cond itions of 1. 0mm o l/L IPTG, 5 h and 30 ℃ induc tion. The expression leve l o f the target fusion pro te in reached 40% of the to tal bac terial pro tein. The resu lts o f SDS- PAGE indicated tha tm o lecularw e ight of the expressed prote in was abou t 22 000 and the expressed pro te in m a inly ex isted in the supe rnatan t after sonication. W estern b lo t ana lysis proved tha t the recom b inant pro tein has good reactive ability aga instm ouse anti-hum an so lub le BAFF IgG. The expression product w as purified by Sephadex G- 75. The pur ified recom bina tion prote in displayed antim icrob ial activ ity obviously.

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Last Update: 2013-05-05