红鲫促性腺激素α亚基的cDNA克隆  

cDNA Cloning of Two Gonadotropin Alpha Subunit from the Red Crucian Carp

作　　者：李建中[1] 陶敏[1] 张轩杰[1] 刘少军[1] 刘筠[1] 

机构地区：[1]湖南师范大学生命科学学院蛋白质化学与发育生物学教育部重点实验室,中国长沙410081

出　　处：《湖南师范大学自然科学学报》2005年第1期62-66,共5页Journal of Natural Science of Hunan Normal University

基　　金：湖南省教育厅资助项目(04C379).

摘　　要：　从红鲫(Redcruciancarp)脑垂体中提取总RNA,根据鱼类促性腺激素(GonadotropinHormone,简称GTH)α亚基的保守性,设计并合成了一对20个碱基长的简并引物,用RT PCR方法扩增并克隆了红鲫GTH α亚基的两种不同的cDNA,分别命名为GTH α1和GTH α2.GTH α1和GTH α2分别由363和366个核苷酸组成,分别编码117和118个氨基酸.这两种类型的cDNA的核苷酸和氨基酸的同源性非常高,分别达到了95.0%和96.6%.但是,它们之间也有4个氨基酸的差别,而且GTH α1有一个三联体核苷酸组成的氨基酸的缺失.聚类分析表明,GTH α亚基在鲤科鱼类中具有高度保守性.Totai RNA was isolated from red crucian carp pituitary gland. To obtain gonadotropin hormone (GTH) α subunits a complete amino acid sequence, a pair of 20-metic degenerate consensus primers were designed, based on the conservative amino acid sequences of GTH-α subunits.The first strand cDNA was synthesized using oligo(dT)18 as a primer, the total RNA as a template, and AMV reverse transcriptase. Two types of cDNA encoding the α subunit of GTH were amplified and cloned by polymerase chain reaction(PCR) for the first strand cDNA of red crucian carp. The red crucian carp GTH-α subunit cDNAs were named as α1 and α2. The nucleotide sequence of GTH-α1 cDNA was 363 bp, encoding 117 amino acids, and that of GTH-α2 was 366 bp,encoding 118 amino acids. These two types of GTH α subunits showed higher homology of 95.0% in the nucleotide and 96.6% in the amino acids. Deduced amino acid sequences represented differences in four residuces between al and α2, including a deletion of an amino acid in α1. The phylogenetic trees of vertebrates glycoprotein hormones (GPHs) α subunits indicated that GTH-α subunits were higher conservative in cyprinids.

关 键 词：促性腺激素 Α亚基 CDNA克隆 红鲫 RT-PCR方法 核苷酸组成 氨基酸 总RNA 简并引物 鲤科鱼类 保守性 脑垂体 同源性 三联体 分析表 碱基 

分 类 号：Q959.468[生物学—动物学] TQ464.7[化学工程—制药化工]