荷包猪SLA-DRB基因cDNA的克隆及分子进化特征分析

doi:10.13560/j.cnki.biotech.bull.1985.2015.12.022

生物技术通报 ›› 2015, Vol. 31 ›› Issue (12): 150-157.doi: 10.13560/j.cnki.biotech.bull.1985.2015.12.022

荷包猪SLA-DRB基因cDNA的克隆及分子进化特征分析

姜平^1,2,额尔敦木图¹,高凤山^1,2

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

收稿日期:2015-04-04 出版日期:2015-12-19 发布日期:2015-12-19
作者简介:姜平，女，硕士，研究方向:动物解剖学与免疫学；E-mail:1224763183 @qq.com
基金资助:
国家自然科学基金项目(31172304)，辽宁省博士启动基金项目(20081078)

The Cloning of cDNA of SLA-DRB from Hebao Pigs and the Analysis of Their Molecular Evolutionary Characteristics

Jiang Ping^1,2, Erdemtu¹, Gao Fengshan^1,2

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

Received:2015-04-04 Published:2015-12-19 Online:2015-12-19

摘要/Abstract

摘要：

旨在研究荷包猪SLA-DRB基因的分子特征，设计引物用RT-PCR扩增3头荷包猪DRB基因cDNA，并克隆至pMD18-T Vector，阳性克隆测序并做序列分析，分别进行同源性、分子进化及主要氨基酸变异位点分析。结果表明，成功从3头荷包猪中扩增得到SLA-DRB基因，分别命名为SLA-DRB-HB01-03，经序列测定后，证实cDNA全长为836 bp，其中1-801为ORF区，共编码266个氨基酸。同源性分析显示，SLA-DRB-HB与其他SLA-DRB等位基因的同源性介于90.3%-99.8%之间。分子进化分析表明，荷包猪SLA-DRB-HB独立分支，与其他等位基因相比较，进化更加原始。氨基酸变异位点和多态性分析结果显示，SLA-DRB-HB本身存在一定的多态性。

关键词: 荷包猪, SLA-DRB, 分子进化, 多态性

Abstract:

In order to study the molecular characteristics of SLA-DRB derived from Hebao pigs, a pair of primers were designed to amplify the cDNA of SLA-DRB from 3 Hebao pigs. Then the amplified cDNA were cloned into pMD18-T vector and the positive clones were sequenced and analyzed by homology comparing, molecular evolution analysis, and comparing and analyzing the key amino acid sites. It was shown that the SLA-DRB alleles were amplified successfully from the tissues of the 3 Hebao pigs, and the alleles were designated as SLA-DRB-HB01, 02, and 03. After sequencing, the results showed that the cDNA of SLA-DRB-HB alleles was 836 bp, and the open reading fragments(ORF)of SLA-DRB-HB locating at sites of 1-801, encoding 266 amino acids. By homology analyzing, the SLA-DRB-HB genes had a homology value of 90.3%-99.8% with other SLA-DRB alleles. The analysis of molecular evolution showed that the SLA-DRB-HB alleles were clustered an independent branch of phylogenetic tree, and SLA-DRB-HB evolved more primarily in contrast to other SLA-DRB alleles. The analysis of the mutated amino acids sites and the polymorphism of SLA-DRB-HB demonstrated that they had polymorphism.

Key words: Hebao pig, SLA-DRB, molecular evolution, polymorphism

姜平,额尔敦木图,高凤山. 荷包猪SLA-DRB基因cDNA的克隆及分子进化特征分析[J]. 生物技术通报, 2015, 31(12): 150-157.

Jiang Ping, Erdemtu, Gao Fengshan. The Cloning of cDNA of SLA-DRB from Hebao Pigs and the Analysis of Their Molecular Evolutionary Characteristics[J]. Biotechnology Bulletin, 2015, 31(12): 150-157.

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荷包猪SLA-DRB基因cDNA的克隆及分子进化特征分析

The Cloning of cDNA of SLA-DRB from Hebao Pigs and the Analysis of Their Molecular Evolutionary Characteristics

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