荷包猪SLA-DRa基因的克隆及序列分析

doi:10.16431/j.cnki.1671-7236.2015.02.008

›› 2015, Vol. 42 ›› Issue (2): 264-269.doi: 10.16431/j.cnki.1671-7236.2015.02.008

荷包猪SLA-DRa基因的克隆及序列分析

姜平^1,2, 高凤山², 额尔敦木图¹

1. 内蒙古农业大学兽医学院, 呼和浩特 010018;
2. 大连大学生命科学与技术学院, 大连 116622

修回日期:2014-10-08 出版日期:2015-02-20 发布日期:2015-02-13
通讯作者: 高凤山, 额尔敦木图 E-mail:gfsh0626@126.com;erdemtu962@sina.com
作者简介:姜平(1971-),女,河北涿鹿人,硕士,研究方向:动物解剖学与免疫学,E-mail:1224763183@qq.com
基金资助:
国家自然科学基金(31172304)

Cloning and Sequences Analysis of the SLA-DRa Gene from Hebao Pig

JIANG Ping^1,2, GAO Feng-shan², Erdemtu¹

1. College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot 010018, China;
2. College of Life Science and Technology, Dalian University, Dalian 116622, China

Revised:2014-10-08 Online:2015-02-20 Published:2015-02-13

摘要/Abstract

摘要： 为研究中国特色品系荷包猪SLA-DRa基因(又称SLA-DRa-HB),本试验设计引物,RT-PCR扩增3个个体荷包猪SLA-DRa全基因编码区,并克隆至pMD18-T载体,转化大肠杆菌JM109感受态细胞,经酶切鉴定后筛选阳性克隆测序,比较分析与其他SLA-DRa等位基因的差异,并绘制分子进化树。结果显示,RT-PCR成功扩增出目的基因条带,大小约800 bp。经克隆测序后分析,SLA-DRa-HB基因全长为779 bp,编码区为1-759,共编码252个氨基酸。序列对比分析结果显示,SLA-DRa-HB的特征性变异集中在135、159、202位点。而穿膜区和胞浆功能区(203-252)变异位点为206、248。分子进化树分析显示,SLA-DRa-HB自成一系,且与其他等位基因的进化关系较近。本研究成功克隆荷包猪SLA-DRa基因,为进一步研究其功能奠定基础。

关键词: 荷包猪; SLA-DRa基因; 克隆; 序列分析

Abstract: In order to study SLA-DRa gene (also named as SLA-DRa-HB) from one of special breed of domestic pig,Hebao,primers were designed to amplify the coding domain of SLA-DRa from three Hebao pigs.Then,the amplified fragments were cloned into pMD18-T vector followed by transforming them into Escherichia coli JM109.After cleaving by the restricted enzymes,the positive clones were selected to be sequenced.The differences between SLA-DRa-HB and other SLA-DRa alleles were analyzed by sequences comparing,and then the phylogenetic tree was constructed.The results showed that the interest of the fragments was successfully amplified by RT-PCR and the molecular weight was about 800 bp.By cloning and sequencing,the whole length of SLA-DRa-HB was 779 bp and the open reading fragments (ORF) of SLA-DRa-HB located at sites of 1 to 759 coding for 252 amino acids.Results of sequences comparision and analysis showed that the characterized amino acids of SLA-DRa-HB mainly focused on sites of 135,159 and 202,while the mutational sites in trans-membrane and cytoplasm were 206 and 248.Analyzing from the phylogenetic tree,it showed that SLA-DRa-HB was clustered into one branch and they were relatively closed to other SLA-DRa alleles.In this research,the SLA-DRa alleles were cloned successfully from Hebao pigs and it would lay a base for study the function of the SLA-DRa genes.

Key words: Hebao pigs; SLA-DRa gene; clone; sequence analysis

中图分类号:

Q786

姜平, 高凤山, 额尔敦木图. 荷包猪SLA-DRa基因的克隆及序列分析[J]. , 2015, 42(2): 264-269.

JIANG Ping, GAO Feng-shan, Erdemtu. Cloning and Sequences Analysis of the SLA-DRa Gene from Hebao Pig[J]. , 2015, 42(2): 264-269.

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荷包猪SLA-DRa基因的克隆及序列分析

Cloning and Sequences Analysis of the SLA-DRa Gene from Hebao Pig

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