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Prokaryotic expression, protein purification and polyclonal antiserum preparation of Beauveria bassiana CDEP2 Gene |
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Abstract: |
Subtilisin-like protease was the most important factor for the insecticidal activity of entomopathogenic fungi. Using the specificity primers, a cDNA encoding the subtilisin-like protease CDEP2 gene was amplified from Beauveria bassiana by RT-PCR. The recombination prokaryotic expression plasmid pET28-CDEP2 was constructed by subcloning the maturation CDEP2 protein gene encoding sequence into the vector pET28a. It was transformed into E.coli BL21 (DE3) and induced to express by IPTG. SDS-PAGE results showed that the His-CDEP2 fusion protein was high express in the form of inclusion body and purified by affinity chromatography. The rabbits were immuned by the purified CDEP2 protein to produce polyclonal antiserum. Western blotting analysis indicated that the antiserum could react with the corresponding protein, and was suitable to be used for further analysis of CDEP2 protein. |
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