来自硕大利什曼原虫阿卜杜的无鞭毛体编码基因的克隆与序列分析。

Cloning and sequence analysis of an amastin coding gene from Leishmania major Abdou.

作者信息

Cheng J, Zhong Y, Liu Y, Yang J

机构信息

Gene Therapy Research Center, Institute of Infectious Diseases, 302 Hospital of PLA, Beijing 100039, China.

出版信息

Chin Med J (Engl). 1999 Aug;112(8):698-700.

PMID:11601274

Abstract

OBJECTIVE

To clone a surface protein encoding gene from Leishmania major Abdou parasites.

METHODS

Using Trypanosotidae cruzi (T. cruzi) amastin DNA sequence as a reference, computer search was done on Genbank and dbEST data bases with BLASTN path. A Leishmania major Abdou DNA library has been established and screened by in situ colony hybridization.

RESULTS

No homology sequence to T. cruzi amastin was found in Genbank, but a 309 nt DNA fragment from Leishmania major (L. major) existed in bEST. Leishmania major Abdou DNA library was screened using specific probes synthesized according to 309 nt DNA sequence of T. cruzi amastin gene, and full-length coding sequence for Leishmania major Abdou amastin was cloned. The coding sequence consisted of 552 nt, and translated into 183 amino acid residues. The homology is 23.5% at amino acid sequence level between Leishmania major Abdou and T. cruzi amastins.

CONCLUSION

We have cloned the full length amastin coding DNA for Leishmania major Abdou.

摘要

目的

从硕大利什曼原虫阿卜杜菌株中克隆一个编码表面蛋白的基因。

方法

以克氏锥虫无鞭毛体DNA序列为参照，通过BLASTN程序在Genbank和dbEST数据库中进行计算机检索。构建了硕大利什曼原虫阿卜杜菌株DNA文库，并通过菌落原位杂交进行筛选。

结果

在Genbank中未发现与克氏锥虫无鞭毛体同源的序列，但在dbEST中存在一段来自硕大利什曼原虫的309 nt DNA片段。利用根据克氏锥虫无鞭毛体基因的309 nt DNA序列合成的特异性探针筛选硕大利什曼原虫阿卜杜菌株DNA文库，克隆出了硕大利什曼原虫阿卜杜菌株无鞭毛体的全长编码序列。该编码序列由552 nt组成，编码183个氨基酸残基。硕大利什曼原虫阿卜杜菌株与克氏锥虫无鞭毛体在氨基酸序列水平上的同源性为23.5%。