@article{ref1,
title="Arabinose-controlled expression of htrA in Shigella flexneri through suicide plasmid-mediated homologous recombination",
journal="Military Medical Sciences",
year="2015",
author="Yang, Jing and Lu, Lilan and Xu, Ping and Liu, Hongsheng and Feng, Erling and Wang, Hengliang and Zhu, Li",
volume="",
number="12",
pages="354-356,402",
abstract="OBJECTIVE To achieve arabinose-controlled expression of HtrA strain and detect the expression of HtrA protein.<br><br>METHODS Arabinose promoter with htrA100 was amplified from pACD-htrA vector by PCR and cloned into pGP704 vector.Then, Shigella flexneri 2a strain 301 was transferred with the recombinant plasmid pGD-htrA and an AraC-expression vector.The expressions of HtrA in whole-cell and periplasmic space were detected by Western blotting.<br><br>RESULTS The suicide plasmid-mediated homologous recombinant vector and the inducible HtrA expression strain were successfully constructed.Without arabinose,HtrA protein was hardly detected ,but in the presense of arabinose , HtrA protein could be detected in whole-cell lysate and in periplasmic space lysate by Western blot.<br><br>CONCLUSION Homologous recombination using suicide plasmid can significantly knock down the expression of HtrA protein.After being induced with arabinose , HtrA protein can be expressed normally.<p /><p>Language: zh</p>",
language="zh",
issn="1674-9960",
doi="10.7644/j.issn.1674-9960.2015.05.008",
url="http://dx.doi.org/10.7644/j.issn.1674-9960.2015.05.008"
}