[Fusion expression, purification and bioactivity detection of XXXX defensinalpha (HDalpha)].

@entity1 @entity27600 @entity8696 @entity3646 @entity214
AIM: To obtain recombinant @entity1 defensin alpha(HDalpha) and detect its biological activity, so as to facilitate further research. METHODS: The HDalpha gene fragment with @entity8696 cleavage site was synthesized, and then cloned into pBV220- @entity3646 vector to construct pBV220- @entity3646 -HDalpha. The constructed vector which was confirmed to be correct by sequencing was transformed into @entity27600 alpha and the @entity3646 -HDalpha fusion protein was expressed under temperature induction. After fusion protein was cleaved to remove @entity3646 by @entity8696 , purification and renaturation was performed. HDalpha's characteristics were identified by @entity214 -PAGE and bioactivity detection. RESULTS: After temperature induction, the expressed fusion protein which accounted for about 20% of total bacterial protein existed mainly in the form of inclusion body. After cleaving by @entity8696 , the purity of obtained HDalpha was about 99.8%. Bacteriostatic test and clone forming test showed that recombinant HDalpha could obviously inhibit the growth of bacteria. CONCLUSION: The recombinant expression plasmid for HDalpha gene has been constructed successfully and obtained engineering bacteria can stably express target protein. Furthermore, techniques of purification and renaturation was set out, which lays the foundation for further functional study and application of HDalpha.
