[克隆中国HIV-1 B、C和E亚型的代表性毒株用于异源双链迁移率分析（HMA）]

[Cloning the representative strains of China HIV-1 subtypes B, C and E for heteroduplex mobility analysis (HMA)].

作者信息

Xiao Y, Yao J, Shao Y

机构信息

National Center for AIDS Prevention and Control, Beijing 100050.

出版信息

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 1999 Mar;13(1):33-6.

PMID:12759949

Abstract

OBJECTIVE

To clone the HIV-1 env gene and establish the HMA reagents suitable for use with the Chinese HIV strains.

METHODS

Using Nest-PCR technique, we amplified env gene from representative strains which contain the most proximate consensus of China HIV-1 subtypes B, C and E. The genes were then cloned into pGEM-Teasy vector for HMA subtyping.

RESULT

The env gene of representative strains of China HIV-1 subtypes B, C and E were amplified and cloned into plasmids. The plasmids were used in HMA and compared with those standard env gene of other countries. The heteroduplex bands obtained with ours env genes were more proximate to homoduplex bands. The homogeneity of the corresponding China HIV-1 subtypes with standard TH14(B), IND868(C), MA959 (C) and TH22(E) are 90.2%(B), 88.3%(C), 83.7(C)% and 92.3%(E) respectively.

CONCLUSION

The HMA reagents we established in this study have higher sensitivity in HMA analysis for China HIV-1 strains when compared with the international HMA reagents. The HMA reagents also provide new reference reagents for UNAIDS HIV isolation network.

摘要

目的

克隆HIV-1 env基因并建立适用于中国HIV毒株的核酸杂交分析（HMA）试剂。

方法

采用巢式聚合酶链反应（Nest-PCR）技术，从含有中国HIV-1 B、C和E亚型最相近共有序列的代表性毒株中扩增env基因。然后将这些基因克隆到pGEM-Teasy载体中用于HMA分型。

结果

扩增并克隆了中国HIV-1 B、C和E亚型代表性毒株的env基因到质粒中。将这些质粒用于HMA，并与其他国家的标准env基因进行比较。用我们的env基因获得的异源双链带与同源双链带更接近。相应的中国HIV-1亚型与标准TH14（B）、IND868（C）、MA959（C）和TH22（E）的同源性分别为90.2%（B）、88.3%（C）、83.7%（C）和92.3%（E）。