荣昌猪胸腺素β15A(TMSB15A)cDNA全长克隆、序列信息及组织表达分析

doi:10.11843/j.issn.0366-6964.2017.09.005

畜牧兽医学报 ›› 2017, Vol. 48 ›› Issue (9): 1602-1610.doi: 10.11843/j.issn.0366-6964.2017.09.005

荣昌猪胸腺素β15A(TMSB15A)cDNA全长克隆、序列信息及组织表达分析

龙熙^1,2,3, 蓝静^1,2,3, 郭宗义^1,2,3, 王金勇^1,2,3, 赵久刚^1,2,3*

1. 重庆市畜牧科学院, 荣昌 402460;
2. 农业部养猪科学重点实验室, 荣昌 402460;
3. 养猪科学重庆市市级重点实验室, 荣昌 402460

收稿日期:2017-03-28 出版日期:2017-09-23 发布日期:2017-09-14
通讯作者: 赵久刚,副研究员,博士,主要从事动物遗传学和免疫学研究,E-mail:zhaojg1978@foxmail.com
作者简介:龙熙(1990-),女,四川资阳人,硕士,助理研究员,主要从事猪遗传育种研究,E-mail:13618288075@163.com
基金资助:
重庆市农发资金项目（13405）；国家自然科学基金项目（31201770）；基本科研业务费项目（2013cstc-jbky-00908）；重庆市畜牧科学院基本科研业务费（16435）

The Full-length Cloning, Sequence Information and Expression Analysis of cDNA of Thymosin β15A (TMSB15A) in Rongchang Pig

LONG Xi^1,2,3, LAN Jing^1,2,3, GUO Zong-yi^1,2,3, WANG Jin-yong^1,2,3, ZHAO Jiu-gang^1,2,3*

1. Chongqing Academy of Animal Science, Rongchang 402460, China;
2. Key Laboratory of Pig Industry Sciences of Ministry of Agriculture, Rongchang 402460, China;
3. Chongqing Key Laboratory of Pig Industry Sciences, Rongchang 402460, China

Received:2017-03-28 Online:2017-09-23 Published:2017-09-14

摘要/Abstract

摘要：

旨在克隆荣昌猪TMSB15A基因全长，检测该基因在荣昌猪各组织中的表达特征，并对其基因的结构与功能进行分析。本研究利用RCAE（Rapid-amplification of cDNA ends）-PCR以及RT-PCR（Real-time quantitative PCR）技术克隆荣昌猪TMSB15A基因mRNA全长序列以及检测该基因在荣昌猪各组织中的表达特征。结果表明，荣昌猪TMSB15A基因全长654 bp，其中CDS区长138 bp，编码45个氨基酸，5'UTR和3'UTR分别长86和430 bp。生物信息学分析表明，TMSB15A蛋白的理论分子量为5.2 ku，有10个带负电的氨基酸，9个带正电的氨基酸，蛋白理论等电点为5.3；预测TMSB15A蛋白无疏水区、信号肽结构、跨膜结构域，其主要在细胞质中发挥生理功能；TMSB15A蛋白的三级结构由α-螺旋和无规则卷曲构成。TMSB15A基因编码的氨基酸序列与人、长臂猿以及黑猩猩的同源性最高，为97.8%，其次是骆驼和兔，为93.3%，与马的同源性最低，为11.1%。TMSB15A在荣昌猪免疫相关组织脾和胸腺中表达量最高（P<0.01），在淋巴结和肺中呈中等丰度表达，在心、肝、肾和肌肉中表达量较低。本试验为研究TMSB15A基因的功能奠定了基础，也为荣昌猪抗病育种研究提供了理论支撑。

关键词: 昌猪, TMSB15A, RACE, 基因克隆, 序列分析, 组织表达

Abstract:

The aim of the present study was to clone the full-length cDNA of TMSB15A, analyse the structure and function of TMSB15A gene and determine its expression profile in Rongchang pig. Using the RCAE (Rapid-amplification of cDNA ends)-PCR and RT-PCR(Real-time quantitative PCR), we cloned the full-length of TMSB15A mRNA and detected the expression profile of TMSB15A gene in Rongchang pig. The results showed that the TMSB15A mRNA of Rongchang pig was 654 bp in length. This sequence consisted of 138 bp of CDS, 86 bp of 5' UTR and 430 bp of 3' UTR. The amino acid sequence deduced from TMSB15A encoded a putative protein consisting of 45 amino acids. Bioinformatics analysis indicated that TMSB15A protein had 10 negatively charged amino acids and 9 positively charged amino acids. The theoretical molecular weight of TMSB15A protein was 5.2 ku and the theoretical isoelectric point was 5.3. It was predicted that no hydrophobic amino acids, signal peptides or transmembrane domain were found in TMSB15A protein. TMSB15A protein might play physiological function in the cytoplasm. The tertiary structure of TMSB15A comprised α-helices and random coils. Rongchang pig TMSB15A had the greatest genetic relationship with TMSB15A of Homo sapiens, Nomascus leucogenys and Pan troglodytes (97.8%), followed by Camelus ferus and Oryctolagus cuniculus (93.3%), and the lowest relationship with Equus caballus(11.1%). The highest expression level of TMSB15A was in the spleen and thymus (P<0.01). Medium expression levels were observed in lymph node and lung. The expression of TMSB15A in heart, liver, kidney and muscle was the lowest. These data provide a basis for investigating the functions of TMSB15A and will inform further research in Rongchang pig resistance breeding.

Key words: Rongchang pig, TMSB 15A, RACE, gene clone, sequence analysis, tissues expression

中图分类号:

S828.2

龙熙, 蓝静, 郭宗义, 王金勇, 赵久刚. 荣昌猪胸腺素β15A(TMSB15A)cDNA全长克隆、序列信息及组织表达分析[J]. 畜牧兽医学报, 2017, 48(9): 1602-1610.

LONG Xi, LAN Jing, GUO Zong-yi, WANG Jin-yong, ZHAO Jiu-gang. The Full-length Cloning, Sequence Information and Expression Analysis of cDNA of Thymosin β15A (TMSB15A) in Rongchang Pig[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(9): 1602-1610.

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荣昌猪胸腺素β15A(TMSB15A)cDNA全长克隆、序列信息及组织表达分析

The Full-length Cloning, Sequence Information and Expression Analysis of cDNA of Thymosin β15A (TMSB15A) in Rongchang Pig

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