Adams S E, Johnson I D, Braddock M, Kingsman A J, Kingsman S M, Edwards R M
Department of Molecular Biology, British Bio-technology Ltd, Cowley, Oxford, UK.
Nucleic Acids Res. 1988 May 25;16(10):4287-98. doi: 10.1093/nar/16.10.4287.
The synthesis of a gene for the HIV TAT protein is described using a novel approach that capitalises on the ability to synthesise oligonucleotides of greater than 100 bp in length. It involves the synthesis of large oligomers covering one strand of the desired gene in its entirety and the use of small complementary bridging and adapter oligonucleotides to direct the assembly and cloning of the large oligomers. After ligation to the cloning vector the partially single stranded intermediate is transformed directly into the recipient bacterial host where the plasmid is repaired. The synthetic tat gene has been expressed in HeLa cells and is shown to trans-activate TAR+ but not TAR- HIV LTR-CAT constructs.
本文描述了一种合成HIV TAT蛋白基因的新方法，该方法利用了合成长度超过100 bp寡核苷酸的能力。它涉及合成覆盖所需基因一条链的全部的大寡聚物，并使用小的互补桥接和衔接子寡核苷酸来指导大寡聚物的组装和克隆。连接到克隆载体后，部分单链中间体直接转化到受体细菌宿主中，在那里修复质粒。合成的tat基因已在HeLa细胞中表达，并显示能反式激活TAR+但不能反式激活TAR-的HIV LTR-CAT构建体。
