Arabinose-controlled expression of htrA in Shigella flexneri through suicide plasmid-mediated homologous recombination / 军事医学
Military Medical Sciences
;
(12): 354-356,402, 2015.
Article
in Chinese
| WPRIM
| ID: wpr-600872
ABSTRACT
Objective To achieve arabinose-controlled expression of HtrA strain and detect the expression of HtrA protein.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 .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 .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 .
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
Military Medical Sciences
Year:
2015
Type:
Article
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