猪多潜能性维持因子5 cDNA全序列的克隆与分析  

Porcine Developmental Pluripotency Associated 5 Gene Complete cDNA Cloning and its Bioinformatics Analysis

作　　者：李勇[1] 王立刚[1] 王立贤[1] 乔丽娟[1] 张龙超 颜华[1] 

机构地区：[1]中国农业科学院北京畜牧兽医研究所,北京100193

出　　处：《农业生物技术学报》2010年第2期285-289,共5页Journal of Agricultural Biotechnology

基　　金：国家十一五科技支撑计划项目(No.2006BAD01A08);国家高技术研究与发展计划(863)(No.2006AA10Z199)共同资助

摘　　要：利用前期筛选出的一些与母猪妊娠相关的有价值的新候选基因,选取其中一个位于网络顶端的EST序列(CK455976),根据此EST序列设计引物,以母猪卵巢为材料提取总RNA,运用RT-PCR和RACE-PCR对其进行cDNA全序列克隆,得到多潜能性维持因子5(developmental pluripotency associated5,DPPA5)基因cDNA全长596bp,提交GenBank数据库,登录号为FJ436413。经序列分析发现,其中5'UTR长67bp,3'UTR长175bp,编码区长354bp,编码117个氨基酸。同源性比对分析发现,该基因核苷酸序列与人、小鼠、猕猴和狗的DPPA5基因同源性分别为83%、70%、83%和80%;编码的氨基酸序列同源性分别为74%、55%、74%和69%。经生物信息学分析,预测出DPPA5的相对分子量约13.60kD,等电点5.24,在约1～10、18～29、38～51、60～66和75～110位之间氨基酸存在亲水性区域,DPPA5蛋白主要存在于细胞膜外侧。An expressed sequence tag（EST, CK455976） on the top of sow gestation relevant gene regulatory networks which was constructed by former reacher in author＇s laboratory was selected to do forward research in this paper.Using the total RNA extracted from sow ovaries and primers designed according to the sequence of this EST, cDNA complete sequence of the developmental pluripotency associated 5 was obtained using RT-PCR and RACE-PC, the length of this sequence was 596 bp and the accession number was FJ36413 in GenBank database.Advanced analysis indicated that the complete sequence contained an untranslated region（located in 1～67 and 422～596） and a coding region（located in 65～421） which coded a polypeptide of 117 amino acids.The predicted similarities nucleotide sequence among pig, human, mouse, Macaca mulatta and Canis familiaris were 83%, 70%, 83% and 80% respectively, and the amino acid sequence were 74%, 55%, 74% and 69% respectively.The relative molecular weight of predicted DPPA5 was 13.60 kD, and its isoelectric point was 5.24.Five hydrophilic regions were located in 1～10、18～29、38～51、60～66 and 75～110 and DPPA5 protein mainly in the outside of the membrane.

关 键 词：猪 多潜能性维持因子5 序列克隆 生物信息学分析 

分 类 号：S814.8[农业科学—畜牧学]