遗传 ›› 2008, Vol. 30 ›› Issue (5): 595-601.doi: 10.3724/SP.J.1005.2008.00595

• 研究报告 • 上一篇    下一篇

人猿超科和旧大陆猴7种高等灵长类FKN全基因序列的测定及进化分析

洪晓武1, 张亚平2, 储以微1, 高海峰1, 蒋正刚1, 熊思东1   

  1. 1. 复旦大学上海医学院免疫学系, 免疫生物学研究所, 上海 200032;
    2. 中国科学院昆明动物研究所细胞与分子进化开放实验室, 昆明 650223

  • 收稿日期:2007-10-23 修回日期:2008-02-18 出版日期:2008-05-10 发布日期:2008-05-10
  • 通讯作者: 熊思东

Complete sequence determination and phylogenetic analysis of FKN among seven higher primates including homonids and Old World Monkeys

HONG Xiao-Wu1, ZHANG Ya-Ping2, CHU Yi-Wei1, GAO Hai-Feng1, JIANG Zheng-Gang1, XIONG Si-Dong1   

  1. 1. Department of Immunology, Shanghai Medical College, Institute for Immunobiology, Fudan University, Shanghai 200032, China;
    2. Laboratory of Cellular and Molecular Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China
  • Received:2007-10-23 Revised:2008-02-18 Online:2008-05-10 Published:2008-05-10
  • Contact: XIONG Si-Dong

摘要:

测定人猿超科(人、黑猩猩、大猩猩、红毛猩猩和长臂猿)和旧大陆猴(猕猴和叶猴)7种高等灵长类FKN全基因序列, 探讨其系统进化分析。用简并引物PCR(Degenerated PCR)法分别扩增FKN的3个外显子, 其产物经琼脂糖凝胶回收、纯化后测序, 然后用BioEdit软件剪切拼接FKN基因全序列, 用DNAStar比对后比较基因和氨基酸序列同源性, Mega软件重构FKN基因进化树, 应用Datamonkey分析FKN的负选择位点。序列分析发现人猿超科较旧大陆猴FKN基因除了有散在的点突变外, 还有一明显的30 bp的核苷酸缺失突变; 人FKN基因序列与黑猩猩、大猩猩、红毛猩猩、长臂猿、猕猴和叶猴的同源性分别是99.2%、98.4%、98.1%、96.5%、95.9%和93.8%, 由此推导的氨基酸序列同源性分别是98.5%、98.0%、97.7%、94.7%、93.7%和90.5%; FKN基因进化树表明人与黑猩猩关系更近, FKN基因进化和通常认为的物种进化一致; Datamonkey分析结果显示FKN存在3个负选择位点53Q、84D、239N。成功获得人、黑猩猩、大猩猩、红毛猩猩、长臂猿、猕猴和叶猴7种高等灵长类物种FKN全基因序列, 为后续探讨FKN在高等灵长类物种进化过程中免疫学功能演变及其结构与功能的关系奠定基础。

关键词: 人猿超科, 旧大陆猴, 测序, 进化分析, FKN

Abstract:

To obtain full-length FKN nucleotide sequences of homonids including human, chimpanzee, gorilla, orangutan and gibbon, and Old World Monkeys including macaque and leaf monkey and make phylogenetic analysis, three exons of FKN were amplified by degenerated PCR using obtained peripheral blood cells DNA as template which was ex-
tracted from homonids and Old World Monkeys. After extracting and purifying from agarose gels, PCR products were se-
quenced and then spliced by using BioEdit. All the FKN sequences were aligned and compared the percent identity by using
DNAStar. The phylogenetic tree was constructed using maximum evolution approach in Mega. The negative selection sites
were analyzed by using Datamonkey. There is an apparent 30 bp nucleotides deletion mutation in homonids FKN compar-
ing to that of Old World Monkeys besides other point mutations. Homology of nucleotide sequence between human and
chimpanzee, gorilla, orangutan, gibbon, macaque and leaf monkey is 99.2%, 98.4%, 98.1%, 96.5%, 95.9% and 93.8%,
respectively. Homology of amino acid sequence of them is 98.5%, 98.0%, 97.7%, 94.7%, 93.7% and 90.5%, respectively.
In the same time, the genealogical relationship of human is a lot closer to chimpanzee than it is to gorilla and other apes. It is generally agreed that the evolution rule of FKN gene is in accord with that of the higher primates. In addition, Datamon-key shows that there are 3 negative selection sites 53Q, 84D and 239N in FKN. The full-length FKN gene of human, chim-panzee, gorilla, orangutan, gibbon, macaque and leaf monkey were sequenced successfully, and the FKN sequences analysis lays the foundation for further studying its evolution in immunological function in higher primates and the relation between its structure and function.