Prokaryotic Expression of BoPGIP1 Gene from Broccoli and Stress Tolerance of E.coli BL21 Expressing BoPGIP1 Gene

doi:10.7668/hbnxb.2015.01.005

Abstract

Abstract: In order to further verify the broccoli BopGIPI genes function, the complete coding sequence of BoPGIP1 (993 bp) for protein expression was amplified by RT-PCR with gene specific primers designed according to multiple clone sites in prokaryotic expression vector pET28a(+) and BoPGIP1 which was obtained in author's previous experiment.The gene was cloned into pET28a(+) vector, and transferred into E.coli strain BL21 (DE3) for heterologous expression after induced by IPTG.SDS-PAGE was used to analyze expression of the target protein.The result showed that there was a specific band at about 37 kDa in size, which was identical with the expected molecular weight of the recombinant protein.Meanwhile, stress tolerance of E.coli BL21 expressing BoPGIP1 gene was analyzed.The results indicated that E.coli line BL21(pET28a- BoPGIP1) had more obvious tolerance to salt, alkali and dry stress than E.coli BL21 (pET28a), these provide preliminary information for the application of BoPGIP1 gene in plant gene engineering.

Key words: Broccoli, BoPGIP1, Prokaryotic expression, Stress resistance

CLC Number:

Q78
S634.03

ZHANG Tao. Prokaryotic Expression of BoPGIP1 Gene from Broccoli and Stress Tolerance of E.coli BL21 Expressing BoPGIP1 Gene[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(1): 24-28. doi: 10.7668/hbnxb.2015.01.005.

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