【摘要】 为了获得高山红景天中红景天苷生物合成关键酶——UDP-糖基转移酶家族基因的全长cDNA序列,采用在线简并引物设计软件结合3′-RACE技术获得两个糖基转移酶基因的3′端部分序列,进一步采用不同的5′-RACE试剂盒扩增两个基因的5′端序列,结果显示利用Takara公司的试剂盒扩增可以得到cDNA全长序列(GenBank登录号为EF508689和EU567325),而采用Invitrogen公司的试剂盒得到的序列不包括全部开放读码框。实验证明,利用Takara公司的SMARTerTM RACE cDNA Amplification Kit克隆获得基因全长序列的比率更高。更多还原

【Abstract】 In order to obtain the critical enzyme for the synthesis of Rhodiola sachalinensis glycoside,two putative UDPglycosyltransferase(UGT) cDNAs(GenBank accession number,EF508689 和 EU567325) were isolated from R.sachalinensi.The primers were designed for 3 ’-rapid amplification of cDNA ends(RACE) based on the consensus hybrid oligonucleotide primers(CODEHOP) strategy by the Block Maker program.A full-length cDNA sequence(Genbank accession numbers:EF508689 and EU567325) was obtained through the amplification using a SMARTerTM RACE cDNA Amplification Kit(TaKaRa,Dalian,China),whereas the sequence obtained using a 5’-RACE system(version 2.0,InvitrogenTM Life Technologies) contained no full-open reading frame.The cloning efficient of two 5’-RACE systems was investigated.Results exhibited that TaKaRa SMARTerTM RACE cDNA Amplification Kit was more efficient in complete cDNA sequence cloning,compared with the InvitrogenTM 5’-RACE system.更多还原