限制性显示技术制备HIV-1B亚型基因片段的研究

Preparation of Gene Fragments from HIV-1 U26942 by Restriction Display

In order to isolate expeditiously the HIV1U26942 DNA fragments for preparation of DNA microarrays, the multiple gene fragments with sizes suitable for DNA microarrays, produced by digesting the dissociated HIV gene with Sau 3AⅠ, were ligated with universal adapters. PCR primers were designed to match the universal adapters (including the restriction site sequence) but with one "nesting" base overhanging at the 3’ end. The PCR reactions that were performed with various single primers or primer combinations were divided into ten subgroups. PCR products were purified and then cloned into the T vectors. The positive clones were propagated and the plasmids were extracted. The target HIV gene fragments were isolated and sequenced, which were correlated precisely with the prediction of restriction analysis. Eighteen gene fragments ranging from 0.1kb to 1kb were prepared for DNA microarray. Restriction display is an effective and rapid method for the isolation of gene

In order to isolate expeditiously the HIV1U26942 DNA fragments for preparation of DNA microarrays, the multiple gene fragments with sizes suitable for DNA microarrays, produced by digesting the dissociated HIV gene with Sau 3AⅠ, were ligated with universal adapters. PCR primers were designed to match the universal adapters (including the restriction site sequence) but with one 'nesting' base overhanging at the 3' end. The PCR reactions that were performed with various single primers or primer combinations were divided into ten subgroups. PCR products were purified and then cloned into the T vectors. The positive clones were propagated and the plasmids were extracted. The target HIV gene fragments were isolated and sequenced, which were correlated precisely with the prediction of restriction analysis. Eighteen gene fragments ranging from 0.1kb to 1kb were prepared for DNA microarray. Restriction display is an effective and rapid method for the isolation of gene fragments