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Updated: Sep 26, 2026
A Restriction Enzyme Based Cloning Method to Assess the In vitro Replication Capacity of HIV-1 Subtype C Gag-MJ4 Chimeric Viruses
Published on: August 31, 2014
[Cloning the representative strains of China HIV-1 subtypes B, C and E for heteroduplex mobility analysis (HMA)]
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.
