人参皂苷生物合成相关新基因GBR6的cDNA克隆及序列分析

Molecular Cloning and Sequence Analysis of GBR6, a Novel Candidate Gene Involved in Ginsenoside Biosynthesis from Panax ginseng

从人参根组织中分离总RNA,使用RT-PCR扩增出人参皂苷生物合成相关基因GBR6开放阅读框(ORF)cDNA片断,并将其定向克隆入原核高效表达载体pQE30,阳性克隆经BamHⅠ和PstⅠ双酶切鉴定、测序验证与已知的GBR6ORF序列完全一致.因而成功构建了pQE30-GBR6ORF融合原核表达载体,为下一步进行GBR6功能表达分析奠定了基础.

The total RNA from Panax ginseng root tissues was isolated. The open reading frame of GBR6, a novel candidate gene involved in ginsenoside biosynthesis was amplified by RT PCR, and was directionally subcloned to pQE30 vector. Restriction enzyme idetification with both BamHⅠand PstⅠand sequence analysis of GBR6 positive recombinants were conducted, subsequently. The result was consistent with the registrated GenBank GBR6 ORF nucleotide seguence size.As a result, a pQE30 GBR6 ORF fusion protein prokaryotic expression vector was successfully constructed, and provided a basis for further prokaryotic expression analysis.