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。
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 SMARTer^TM RACE cDNA Amplification Kit（TaKaRa,Dalian,China）,whereas the sequence obtained using a 5＇-RACE system（version 2.0,Invitrogen^TM Life Technologies） contained no full-open reading frame.The cloning efficient of two 5＇-RACE systems was investigated.Results exhibited that TaKaRa SMARTer^TM RACE cDNA Amplification Kit was more efficient in complete cDNA sequence cloning,compared with the Invitrogen^TM 5＇-RACE system.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2010年第21期244-247,共4页
Food Science
基金
国家自然科学基金项目(30900112)
