中国寄生虫学与寄生虫病杂志 ›› 2009, Vol. 27 ›› Issue (4): 1-296.

• 论著 •    下一篇

湖北钉螺线粒体基因组全序列测定研究

李石柱1,王艺秀1,2,刘琴1,吕山1,王强1,吴缨1,张仪1,周晓农1 *   

  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2009-08-30 发布日期:2009-08-30

Complete Mitochondrial Genome Sequence of Oncomelania hupensis (Gastropoda: Rissooidea)

LI Shi-zhu1,WANG Yi-xiu1,2,LIU Qin1,LV Shan1,WANG Qiang1,WU Ying1,ZHANG Yi1,ZHOU Xiao-nong1 *
  

  • Received:1900-01-01 Revised:1900-01-01 Online:2009-08-30 Published:2009-08-30

摘要:

目的 测定并分析湖北钉螺(Oncomelania hupensis)线粒体基因组全序列。 方法 利用特异引物和通用引物分别扩增湖北钉螺线粒体细胞色素C氧化酶Ⅰ(COⅠ)、细胞色素b(Cytb)、16SrRNA(16S)和细胞色素C氧化酶Ⅲ(COⅢ)基因片段,在此基础上利用长PCR技术扩增上述4个基因间的长片段,纯化克隆后采用引物步移法测序。 结果 湖北钉螺线粒体基因组全序列为15 182 bp(GenBank登记号为FJ997214),为闭合环状分子,A+T含量为67.3%;包括13个蛋白基因、22个tRNA基因、2个RNA基因和一段72 bp的A+T富集区;蛋白质编码基因均以ATG为启动子,除呼吸链NADH脱氢酶的第一亚单位(ND1)基因以潜在的T作为终止密码子外,其余基因均以典型的TAA或TAG为终止子;基因重叠区有2处,分别为4 bp和7 bp;基因间隔区共21处合计145 bp,长度范围为1~30 bp;22个tRNA中,除2个tRNASer和tRNAGln、tRNAIle以外均能形成典型的二级结构。 结论 获得了湖北钉螺的线粒体基因组全序列。

关键词: 湖北钉螺, 线粒体基因组, 引物步移法, 序列分析

Abstract:

Objective To sequence and analyze the complete nucleotide sequence of the mitochondrial genome of Oncomelania hupensis. Methods Four long fragments were amplified by long PCR using the primers designed based on mtDNA-COⅠ, Cytb, 16S rRNA and COⅢ gene sequences, and sequenced by conserved primer-walking. Rusults The mitochondrial genome (GenBank accession no. FJ997214) was a circular molecule of 15 182 bp with a total A+T content of 67.32%, and contained 13 protein-coding genes, 2 ribosomal RNA genes, 22 tRNA genes, and an A+T-rich region of 72 bp. All 13 protein-coding genes of the O. hupensis mtDNA used ATG as start codon. Canonical TAA and TAG termination codons were found in 12 protein-coding genes, and the remaining one (ND1) had an incomplete termination codon (T). Two short gene overlaps were found with a length of 4 bp and 7 bp, respectively. The length of 21 total intergenic region of mtDNA was 145 bp ranging from 1-30 bp. A total of 22 transferring RNA were found, all of which were typical cloverleaf structure except for two tRNASer, one tRNAGln and one tRNAIle. Conclusion The complete sequence of O. hupensis mitochondrial genome has been determined.

Key words: Oncomelania hupensis, Mitochondrial genome, Primer-walking, Sequence analysis