中国寄生虫学与寄生虫病杂志 ›› 2000, Vol. 18 ›› Issue (6): 1-324.

• 论著 •    下一篇

我国内脏利什曼病山丘疫区与平原疫区利什曼原虫SSUrDNA多变区序列分析

卜玲毅1;胡孝素1;敬保迁2;易桃林1
  

  1. 华西医科大学寄生虫学研究室,成都 610044;川北医学院微生物学教研室,南充 637000
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2000-12-30 发布日期:2000-12-30

SEQUENCE ANALYSIS OF SSU rDNA VARIABLE REGIONS OF LEISHMANIAISOLATES FROM HILLY FOCI AND PLAIN FOCI OF CHINA

BU Ling-yi 1;HU Xiao-su 1;JING Bao-qian 2;YI Tao-lin 1
  

  1. 1 Department of Parasitology;West China University of Medical Sciences;Chengdu 610041; 2 Department of Microbiology;North Sichuan Medical College;Nanchong 637000
  • Received:1900-01-01 Revised:1900-01-01 Online:2000-12-30 Published:2000-12-30

摘要:   [目的 ]分析我国内脏利什曼病 (VL)山丘疫区与平原疫区利什曼原虫 (L .d )分离株小亚基核糖体DNA (SSUrDNA)多变区的序列差异。 [方法 ]nDNA进行PCR扩增 ,将扩增出的SSUrDNA基因的特异片段克隆于 pGEMR TEasyVector上 ,采用通用引物M 13进行PCR扩增 ,全自动测序仪测序。 [结果 ]序列分析显示 ,扩增的 5株利什曼原虫SSUrDNA序列大小均为 392bp ;序列差异均发生在两个独特序列区 (UQ Ⅰ和UQ Ⅱ ) ;山丘疫区L .d甘肃分离株和四川分离株均在UQ Ⅱ区上有 2个相同的碱基突变 ,L .d甘肃分离株UQ Ⅰ区上有 1个碱基突变 ;无移码突变。 [结论 ]山丘疫区分离株与平原疫区分离株之间的碱基序列有差异 ,平原疫区L .d山东分离株与婴儿利什曼原虫 (L .infantum)的碱基序列相同

关键词: 利什曼原虫, 小亚基核糖体核酸基因, 聚合酶链反应, 克隆, 序列分析, 基因变异

Abstract:

 Objective] To analyze the sequence difference of the SSU rDNA variable regions of Leishmania isolates from hilly foci and plain foci of China. [Methods] Specific SSU rDNA fragments from nuclear DNA of five Leishmania species and isolates were amplified by PCR. The amplified DNA fragments were cloned into pGEM R-T Easy vector. The specific fragments were sequenced by the automated DNA sequencer. [Results] Sequence analysis showed that the amplified DNA fragments of five Leishmania species and isolates were all 392 bp in length, point mutations were located in the two unique sequence (UQ-Ⅰ and UQ-Ⅱ); L.d.SC10 and L.d.GS7 had two same point mutations in UQ-Ⅱ, only L.d.GS7 had one in UQ-Ⅰ; no insertion/deletion. [Conclusion] Sequence difference of the SSU rDNA variable region existed between Leishmania isolates from hilly foci and plain foci; The sequences of the SSU rDNA variable regions of L.d. SD2 isolate and L.infantum were identical.

Key words: Leishmania, small subunit ribosomal DNA, polymerase chain reaction, cloning, sequence analysis, genetic variation