培养的人类细胞中人类免疫缺陷病毒DNA之间的同源重组：影响重组的因素分析。

Homologous recombination between human immunodeficiency viral DNAs in cultured human cells: analysis of the factors influencing recombination.

作者信息

Kalyanaraman S, Jannoun-Nasr R, York D, Luciw P A, Robinson R, Srinivasan A

机构信息

Retrovirus Diseases Branch Centers for Disease Control, Atlanta, Georgia 30333.

出版信息

Biochem Biophys Res Commun. 1988 Dec 30;157(3):1051-60. doi: 10.1016/s0006-291x(88)80981-1.

DOI:10.1016/s0006-291x(88)80981-1

PMID:2462865

Abstract

Recombination between HIV DNAs was analyzed using DNA transfection in cell cultures and the optimal conditions for efficient recombination were determined. Recombinant plasmid DNA substrates were constructed from HIV proviral DNAs and the success of recombination was measured by the production of viable hybrid virus. The process of recombination between HIV DNAs was shown to be i) dependent on homology between the truncated HIV DNAs and ii) maximum with concentrations of the truncated DNAs 3ug and above. HIV isolates with heterogeneity in their primary sequence, thus offer an ideal system for the analysis of the requirement of homologous recombination. In addition, recombination methodology would be useful for generating hybrid HIVs for the analysis of specific viral gene functions.

摘要

利用细胞培养中的DNA转染分析HIV DNA之间的重组，并确定有效重组的最佳条件。从HIV前病毒DNA构建重组质粒DNA底物，并通过产生有活力的杂交病毒来衡量重组的成功与否。HIV DNA之间的重组过程被证明为：i）依赖于截短的HIV DNA之间的同源性；ii）当截短DNA的浓度为3μg及以上时重组效率最高。在其一级序列中具有异质性的HIV分离株，因此为分析同源重组的要求提供了一个理想的系统。此外，重组方法将有助于产生杂交HIV，用于分析特定病毒基因的功能。