猪TPST1基因的克隆、序列分析及真核表达载体的构建  

Cloning,sequence characterization and construction of eukaryotic cell expression vector of TPST1 gene in pig

作　　者：别墅[1] 孙洪涛[1] 张冬杰[2] 汪亮[2] 刘娣 

机构地区：[1]东北农业大学动物科学技术学院,哈尔滨150030 [2]黑龙江省农业科学院,哈尔滨15008

出　　处：《东北农业大学学报》2011年第9期12-16,共5页Journal of Northeast Agricultural University

基　　金：国家"十一五"科技支撑计划重点项目(2008BADB2B02-06)

摘　　要：酪氨酸硫化转移酶(Tyrosylrotein sulfotransferase,TPST)在酪氨酸硫化修饰过程中起调节作用。试验参照人、鼠、牛等动物的TPST1 mRNA全长序列设计引物,通过电子克隆结合RT-RCR的方法首次克隆获得包括完整CDS区的猪TPST1基因cDNA序列(HQ439987),分析其核苷酸序列及其编码蛋白质分子结构特征。结果表明,猪TPST1序列全长1 543 bp,包含1 113 bp的完整开放阅读框,编码370个氨基酸。多序列比对和系统进化分析表明,该基因编码蛋白质与牛(97.6%)、小鼠(96.8%)、大鼠(97%)、人(97.03%)等物种TPST1蛋白质具有较高的一致性。蛋白质序列预测分析发现,猪TPST1氨基酸序列理论等电点(pI)为9.0,相对分子质量(MW)为42.13 ku。CTyrosine O-sulfation is a common posttranslational modification of proteins which is mediated tyrosylprotein sulfotransferase.In this study,Porcine TPST1 cDNA was obtained by reverse transcription polymerase chain reaction（RT-PCR）,using in silico cloning strategy based on porcine ESTs database,the nucleotide and amino acid sequences were characterized with bioinformatics methods.The length of the cloned porcine TPST1 cDNA was 1 543 bp including a 1 113 bp entire open reading frame（ORF）,which encoded a 370 amino acids protein.Multiple sequences alignment and phylogeny analysis indicated that porcine TPST1 protein shared high similarity with that of other species,such as Bos taurus（97.6%）,Musmusculus（96.8%）,Rattus norvegicus（97%） and Homo sapiens.Protein sequence analysis showed that the deduced porcine TPST1 protein sequence（pI=9.0,MW= 42.13 ku）.This study had completed the clone TPST1 of min pig and construction of eukaryotic cell expression vector of pcDNA3.1（＋）-TPST1,and providing the theoretical basis for the expression and function research of TPST1 gene in cell.

关 键 词：民猪 TPST1基因 序列分析 载体构建 

分 类 号：S828[农业科学—畜牧学]