鸭CD8α基因克隆及不同禽类分子进化分析

畜牧兽医学报

鸭CD8α基因克隆及不同禽类分子进化分析

徐琪¹，黄正洋¹，孙志明²，陈阳¹，赵文明¹，张扬¹，李秀¹，段修军³，陈国宏^1*

（1. 江苏省动物遗传繁育与分子设计重点实验室，扬州 225009； 2. 江苏省无锡市动物园，无锡 214151；3. 国家水禽种质资源基因库，泰州 225300）

收稿日期:2012-06-29 出版日期:2012-12-26 发布日期:2012-12-26
通讯作者: 陈国宏，E-mail:ghchen@yzu.edu.cn
作者简介:徐琪（1978-），男，江苏如东人，博士，主要从事家禽遗传育种研究，E-mail: xuqi@yzu.edu.cn
基金资助:
国家自然科学基金项目（31101704）；现代农业产业技术体系建设专项资金（CARS-43-3）；江苏高校优势学科建设工程资助项目（苏政办发【2011】137号）

Molecular Cloning and Evolution of CD8α Gene in Duck Populations

XU Qi¹, HUANG Zheng-yang¹, SUN Zhi-ming², CHEN Yang¹, ZHAO Wen-ming¹, ZHANG Yang¹， LI Xiu¹, DUAN Xiu-jun³, CHEN Gu-hong^1*

(1. Jiangsu Key Laboratory of Animal Genetics ＆ Breeding and Molecular Design, Yangzhou 225009, China； 2. Jiangsu Wuxi Zoo, Wuxi 214151, China； 3. National Waterfowl Germplasm Resorurse, Taizhou 225300, China)

Received:2012-06-29 Online:2012-12-26 Published:2012-12-26

摘要/Abstract

摘要：

采用RT-PCR和LD-PCR技术克隆了5个鸭种（金定鸭、樱桃谷北京鸭、番鸭、绿头鸭和斑嘴鸭）CD8α基因编码区序列，结合GenBank中已有禽类（红色原鸡、北京油鸡、斑胸草雀和火鸡）的CD8α序列，采用Jmodeltest最适核苷酸替代模型筛查，使用DAMBE V4.5.2对核苷酸替换(转换/颠换)的饱和性进行检测；并采用最大似然法检测各位点承受的选择压力。测序结果表明，不同鸭种间仅发现少数点突变，其编码区序列相当保守；AIC信息量准则检验结果发现9种禽类CD8α基因的最优核苷酸替代模型为“GTR+G”，核苷酸替换的饱和性检测结果表明序列的核苷酸替换并未饱和；应用 MEGA 5.0 构建了禽类CD8α基因的系统发育关系树，不同禽类CD8α基因大致划分为 3类；“位点-特异”模型检测到7个受到正选择作用的位点，其中4个位于功能重要的区域。本研究成功地克隆了鸭CD8α基因编码区序列，并发现不同禽类CD8α基因在分子进化中是保守的，研究结果为进一步了解禽类CD8α基因进化历程、结构与功能变异提供理论依据。

Abstract:

The coding sequence of CD8α was cloned by RT-PCR and LD-PCR in five duck populations (Jinding Duck, Cherry Valley duck, muscovy, mallard and spot-billed duck). The 9 sequences （another 4 sequences from GenBank） were analyzed by online software of bioinformatics(the best nucleotide substitution model) was selected by Jmodeltest, the nucleotide substitution saturation was assessed with DAMBE, and the selective pressure on amino acid sites was tested by PAML. The results of sequencing showed the coding sequence of CD8α was conservative except a few point mutations. The model “GTR+G” was selected as the best nucleotide substitution model, the sequences were experienced full substitution unsaturation. The phylogenetic trees of CD8α among different fowl were generated by MEGA 5.0, and the 9 fowl were divided into 3 parts according to CD8α sequences. The 7 amino acid sites undergoing positive selection were detected, 4 of them were in the important function regions by the site-specific model. In summary, the coding sequence of CD8α was cloned successfully，and it was conservative in molecular evolution, which may be helpful for studying evolution, structure and function of CD8α among fowl.

中图分类号:

S852.4

徐琪，黄正洋，孙志明，陈阳，赵文明，张扬，李秀，段修军，陈国宏. 鸭CD8α基因克隆及不同禽类分子进化分析[J]. 畜牧兽医学报, doi: .

XU Qi, HUANG Zheng-yang, SUN Zhi-ming, CHEN Yang, ZHAO Wen-ming, ZHANG Yang，LI Xiu, DUAN Xiu-jun, CHEN Gu-hong. Molecular Cloning and Evolution of CD8α Gene in Duck Populations[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, doi: .

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