摘要
[目的 ]分析新疆皮肤利什曼病病原体SSUrRNA基因序列 ,为该病原体种株鉴定分析提供实验依据。[方法 ]应用利什曼原虫特异引物R2 2 2及R333,PCR扩增SSUrRNA基因多变区的片段 ,克隆于T Easyvector,并以双脱氧链末端终止法进行序列测定 ,用DNASIS软件和GenBank微机联网检索分析所测XJCLP、L infantum及L tropicaSSUrRNA基因序列。[结果 ]XJCLP、L infantum及L tropicaSSUrRNA基因插入片段长度均为 394bp。XJCLP与L tropicaSSUrRNA基因 394bp序列存在 3个点突变 ,两者有 391个碱基相同 ,相似性为 99 2 %。XJCLP与L infantumSSUrRNA基因序列存在 3个点突变及 1个移码突变 ,两者有 390个碱基相同 ,类似性为99 0 %。L infantum与L tropica的SSUrRNA基因序列存在 1个移码突变 ,相似性为 99 7%。XJCLP、L infantum及L tropicaSSUrRNA基因序列一级结构与RNA二级结构均不同。经用GenBank检索本次所测XJCLP、L infantum及L tropicaSSUrRNA基因序列 ,该GenBank内尚无同类 394bp的SSUrRNA基因序列。[结论 ]XJCLP、L infantum及L tropica的SSUrRNA基因 394bp序列一级结构 ,二级结构存在差异。
Objective] By sequencing of SSU rRNA gene cloning from Xinjiang cutaneous leishmaniasis pathogen (XJCLP) to provide evidence for identification of the pathogen. [Methods] By PCR assay with primers R222 and R333, the specific fragment had been produced from SSU rRNA gene of XJCLP , L infantum, L tropica and cloned into pGEM ○[KG-6/7]R T Easy vector .The clones were sequenced by the Sanger dideoxy mediated chain termination method, analysis of SSU rRNA gene sequences from XJCLP, L tropica, L infantum with DNASIS. [Results] Sequence analysis showed that the specific fragment of SSU rRNAgene from XJCLP, L infantum,L tropica , were all 394 bp in length. There were 391 bases identical and three point mutations between the sequences of XJCLP and L tropica , the similarity being 99 2%; 390 bases identical and three point mutations and one insertion /deletion between the sequences of XJCLP and L infantum , the similarity being 99 0%. One insertion/deletion between the sequences of L tropica and L infantum , the similarity being 99 7%. The primary and secondary structures of SSU rRNA gene from XJCLP differed from those of L infantum and L tropica .A retrieval from GenBank confirmed that these 394 bp sequence are new gene sequences. [Conclusion]The primary and secondary structures of SSU rRNA gene from XJCLP, L infantum , L tropica were different. 394 bp sequence from SSU rRNA gene of XJCLP is a new gene sequence.
出处
《中国寄生虫学与寄生虫病杂志》
CAS
CSCD
北大核心
2000年第5期260-262,共3页
Chinese Journal of Parasitology and Parasitic Diseases
基金
国家自然科学基金!资助项目 (No .39970 6 6 7)
广东省自然科学基金!资助项目 (No . 9940 5 2 )&&
