Cloning and prokaryotic expression of riboflavin kinase gene in Panax ginseng / 中草药

ABSTRACT

Objective To clone the riboflavin kinase gene of Panax ginseng and perform bioinformatics analysis to construct the prokaryotic expression vector and induce its expression in Escherichia coli. Methods The sequences of comp61599_c0_seq12 were screened from transcriptome database in P. ginseng, and software and online resources were used for its bioinformatics analysis. The 3’ end of sequence was lost and the full-length of 3’ end sequence was obtained by 3’ RACE technology. The complete sequences of cDNA of riboflavin kinase in ginseng was obtained by PCR amplification, and the prokaryotic expression vector of pET-32a-PgRFK was constructed and transformed into E. coli BL-21 for inducing expression. Results A riboflavin kinase gene with length of 1 200 bp was successfully cloned from P. ginseng which encoded 399 amino acid and its prokaryotic expression vector was successfully constructed. The molecular weight of SDS-PAGE electrophoresis results was consistent with prediction. Conclusion The PgRFK gene was successfully cloned and expressed in E. coli.