银杏类黄酮糖基转移酶基因片段克隆与分析

根据GenBank中其他植物的类黄酮糖基转移酶基因(UFGT)的保守区设计简并引物,采用RT-PCR技术从银杏中扩增出UFGT基因的部分保守序列,并利用3′-RACE PCR方法,得到了包括3′-末端在内的银杏UFGT基因片段,命名为GbUFGT-3。结果表明:该片段长度为1 064bp,其中3′-UTR长度为314bp、编码区长度为750bp,可编码249个氨基酸,该氨基酸序列具有完整的UFGT特征序列PSPG-box;BLASTp分析及系统进化树分析表明GbUFGT-3确为银杏UFGT基因的序列片段;半定量RT-PCR结果表明,GbUFGT-3是非诱导表达基因,在银杏叶片的整个发育期均有较高水平的表达,不同发育期表达量无差异。GbUFGT-3已被GenBank收录,登录序列号为JN640564.1。

Molecular cloning and analysis of flavonoid glycosyltransferase gene fragment from Ginkgo biloba

According to plant UFGT genes in GenBank,a 1 064 bp long cDNA fragment containing a 314 bp 3′-UTR and a 750 bp coding region,named GbUFGT-3 was obtained from leaves of Ginkgo biloba using degenerate primer PCR and 3′-RACE protocol,which had been embodied in GenBank database with an accession No.of JN640564.1.The cloned GbUFGT-3 cDNA fragment could encode 249 amino acids,which containes the conserved PSPG-box.BLASTp and phylogenetic tree analysis showed that GbUFGT-3 was partial of UFGT gene from Ginkgo biloba.Semi-quantitative RT-PCR analysis revealed that GbUFGT-3 was an uninduced gene,which could be highly expressed in different developmental stages of Ginkgo biloba leaf with the same expression level.