Construction of prokaryotic expression vector and expression of GbMYBFL protein

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Research Paper 01/08/2018
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Construction of prokaryotic expression vector and expression of GbMYBFL protein

Liangqiong Ma, Weiwei Zhang, Feng Xu, Yongling Liao
Int. J. Biosci.13( 2), 105-110, August 2018.
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Flavonoids are one of the main secondary metabolites of Ginkgo biloba, and MYB is a transcription factor involved in regulation of flavonoid synthesis. In order to construct the prokaryotic expression vector of GbMYBFL from Ginkgo biloba and express the target protein, a pair of primers were designed with Bam H I and EcoR I restriction sites to amplify the ORF region of Ginkgo GbMYBFL gene by PCR. The PCR product was digested by BamH I and EcoR I, and inserted into the prokaryotic expression vector pET32a. Then the recombinant plasmid was transformed into E.coli BL21 (DE3). After induction and expression, the protein was detected by SDS-PAGE. The results showed that the prokaryotic expression recombinant plasmid pET32a-GbMYBFL was successfully constructed. The recombinant plasmid was induced by IPTG in E. coli BL21 (DE3), and the size of the protein electrophoresis band is about 48 kDa, which was consistent with the molecular weight of protein predicted by bioinformatics tools. This study laid the foundation for the functional research of GbMYBFL in Ginkgo biloba.


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