非洲菊查尔酮合酶基因的克隆、序列分析及在大肠杆菌中的表达

Chalcone Synthase Gene Cloning in Gerbera hybrida and Expression in E.coli

摘要:

利用RT-PCR方法,从非洲菊(Gerberahybrida)花瓣的cDNA中克隆到了查尔酮合酶(ChalconeSynthase,CHS)基因CHS,进行了序列分析。结果表明,克隆到的CHS基因全长为1197bps,编码一个由398个氨基酸残基组成的多肽,与Helariutta等发表的非洲菊查尔酮合酶CHS1基因的cDNA序列的CHS基因同源性高达99%。进一步将该基因克隆到表达载体pET32a上,经IPTG诱导表达,得到高效表达的融合蛋白。

Abstract:

Chalcone synthase (CHS) gene was cloned from the petals of Gerbera hybrida using RT-PCR method in our experiment. The sequence of the cDNA coding region was 1 197 bps, encoding a protein of 398 amino acids sharing 99% identical to CHS1 cDNA reported by Helariutta. The gene was fused in expression vector pET32a and was efficiently expressed by transforming into E. coli BL21(DE3) with IPTG induction.