蕙兰GAPDH基因的全长克隆及生物信息学分析  

Clone and Bioinformatics Analysis of GAPDH From ORCHID( Cymbidium faberi)

作　　者：田云芳[1] 蒋素华[1] 袁秀云[1] 崔波[1] 

机构地区：[1]郑州师范学院生物研究所,河南郑州450044

出　　处：《江西农业大学学报》2013年第4期870-877,共8页Acta Agriculturae Universitatis Jiangxiensis

基　　金：河南省重点科技攻关项目(092102110128);郑州市科技计划项目(112PPTGY250-3)

摘　　要：根据GenBank中已登陆的GAPDH基因的同源核苷酸保守序列,设计简并引物,采用RT-PCR的方法得到5个875 bp的片段,分别命名为CfGAPDH1-CfGAPDH5,GenBank登录号分别为JN177722-JN177726。同时结合RACE方法得到蕙兰GAPDH基因cDNA全长,命名为CfGAPDH(JX560732),该序列cDNA全长为1 496 bp,具有完整的开放阅读框(ORF,173-1 195 bp),编码340个氨基酸,该基因编码的氨基酸序列与其他植物的GAPDH蛋白有较高的同源性(80%以上)。系统进化分析表明蕙兰CfGAPDH基因核苷酸序列与墨兰GAPDH1、GAPDH2和春兰GAPDH1的同源性最高(99%),所编码的氨基酸序列与墨兰GAPDH2的同源性达100%。生物信息学分析推测,该基因编码的蛋白残基主要有两个卷曲螺旋区域,分别位于25和250位点附近,属于不稳定蛋白,亲水性较强,初步推断CfGAPDH是一个胞质型GAPDH,二级结构中的β转角所占比例较高(41.2%),三级结构与春兰和小麦非常相似。With primers designed based on the sequences of GAPDH genes from NCBI, the full - length cDNAs of GAPDH genes were cloned from Cymbidium faberi by RT - PCR combined with RACE. The newly cloned GAPDH gene was designated as CfGAPDH （JX560732）. Sequence analysis shows that CfGAPDH is 1 496 bp in length,contains a single open reading frame from 173 bp to 1 195 bp,encodes proteins with 340 a- mino acids. CfGAPDH and Cymbidium sinense GAPDH2 （ JQ796075 ） are more similar having 100% identical amino acids. Bioinformaties analysis showed that the deduced protein is a putative unstable hydrophilie protein. The secondary structure of CfGAPDH consists mainly of loops （41.2%）. The homology modeling of the GAP- DH in Cymbidiumfaberi based on the 3D structure is similar to that of homologues in Cymbidium goeringii and Triticum aestivum,which firms that the polypeptides are highly conserved.

关 键 词：蕙兰 GAPDH RACE 生物信息学 

分 类 号：S682.31[农业科学—观赏园艺]