中国寄生虫学与寄生虫病杂志

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中华按蚊丝氨酸蛋白酶抑制因子14（SRPN14）基因的多态性和进化研究

冯欣宇1,4，马雅军1 *，徐建农2，梁江涛3，夏爱3   

  1. 1第二军医大学热带医学与公共卫生系热带传染病学教研室，上海 200433；2 美国新墨西哥州立大学生物系，拉斯克鲁塞斯 88003；3 南京农业大学昆虫学系，南京 210095；4 中国疾病预防控制中心寄生虫病预防控制所，卫生部寄生虫病原与媒介生物学重点实验室，世界卫生组织疟疾、血吸虫病和丝虫病合作中心，上海 200025
  • 出版日期:2015-08-30 发布日期:2015-09-10

Genetic Polymophism and Evolution of SRPN14 Gene in Anopheles sinensis（Diptera ∶ Culicidae）

FENG Xin-yu1,4, MA Ya-jun1 *, XU Jian-nong2, LIANG Jiang-tao3, XIA Ai3   

  1. 1 Department of Tropical Infectious Diseases, Faculty of Tropical Medicine and Public Health, The Second Military Medical University, Shanghai 200433, China; 2 Department of Biology, New Mexico State University, Las Cruces NM 88003, USA; 3 Entomology Department, Nanjing Agricultural University, Nanjing 210095, China; 4 National Institute of Parasitic Disease, Chinese Center for Disease Control and Prevention, Shanghai 200025, China
  • Online:2015-08-30 Published:2015-09-10

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摘要：

目的  鉴定和定位中华按蚊丝氨酸蛋白酶抑制因子14（SRPN14）基因，分析其在不同群体中的多态性，以及进化中的选择压力。  方法  依据中华按蚊基因组测序数据，设计引物，建立扩增SRPN14基因部分片段的体系，荧光原位杂交进行染色体定位。测定采自我国12省（市）18个采集地的中华按蚊群体的SRPN14基因部分片段序列，计算其在群体内与群体间的遗传差异，以及适应性进化的选择压力。  结果  本研究获得的中华按蚊SRPN14基因部分序列长度为429 bp，与冈比亚按蚊SRPN14基因的对应位置在核苷酸和氨基酸水平的序列一致性分别为77%和88%，位于唾腺染色体的2L：23C亚区。采自我国13个群体411个个体中，SRPN14的等位基因数目共204个，其中51个在群体间共享，占25.00%。13个群体的中华按蚊SRPN14等位基因数目范围为11（辽宁群体LN）~33（重庆群体CQ），核苷酸多态性为0.008（LN）~0.024（海南群体HAN）。AMOVA分析结果显示，群体内变异显著大于群体间，群体内变异占总变异的95.79%，群体间差异小，遗传分化指数（FST）值为0.042。中华按蚊各群体SRPN14基因核苷酸同义替换位点数明显多于非同义替换位点数，ω均小于1。  结论  SRPN14基因在中华按蚊群体中存在一定的多态性，其进化受到负选择压力。

关键词: 中华按蚊, 丝氨酸蛋白酶抑制因子14, 多态性, 进化

Abstract:

Objective  To identify and locate the serine protease inhibitor 14（SRPN14） gene of Anopheles sinensis, and analyze its genetic polymorphism among populations as well as the selective pressure during evolution.  Methods  Primers were designed based on the genomic sequencing data of An. sinensis, and PCR amplification system for the SRPN14 gene was established. The chromosomal location of SRPN14 gene was determined by fluorescence in situ hybridization. The SRPN14 gene of An. sinensis populations collected from 18 sampling sites in 12 provinces（municipality）was sequenced, its genetic variations within and among populations calculated, and the selective pressure during adaptive evolution evaluated.  Results  The amplified part of the SRPN14 gene of An. sinensis was 429 bp in length, and had 77%（nt） and 88%（aa）similarities with An. gambiae. The SRPN14 gene located on 2L: 23C of salivary gland chromosomes of An. sinensis. The sequences of 411 individuals from 13 An. sinensis populations were analyzed. In the 411 individuals, the total number of alleles of the SRPN14 gene was 204, among which 51（25.00%） showed inter-population consistency. The ranges of SRPN14 allel number and genetic polymorphism were from 11 （Liaoning） to 33 （Chongqing）, and from 0.008 （Liaoning） to 0.024 （Hainan）, respectively. AMOVA result showed that genetic divergence within populations was significantly higher than that among populations; variation within populations was 95.79% of the total variation. The genetic divergence among populations was small, with FST value of 0.042. The number of synonymous substitutions in SRPN14 was significantly higher than that of non-synonymous substitutions sites, and ω was less than 1 in all populations.  Conclusion  Genetic polymorphism occurs in SRPN14 gene of An. sinensis populations, and its evolution is under the negative selective pressure.

Key words: Anopheles sinensis, SRPN14, Genetic polymorphism, Evolution

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