SYNTHESIS OF A GENE FOR THE HIV TRANSACTIVATOR PROTEIN TAT BY A NOVEL SINGLE STRANDED APPROACH INVOLVING INVIVO GAP REPAIR
SYNTHESIS OF A GENE FOR THE HIV TRANSACTIVATOR PROTEIN TAT BY A NOVEL SINGLE STRANDED APPROACH INVOLVING INVIVO GAP REPAIR
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DOI：
10.1093/nar/16.10.4287
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发表时间：
1988-05-25
影响因子：
14.9
通讯作者：
EDWARDS, RM
中科院分区：
文献类型：
--
作者：
ADAMS, SE;JOHNSON, ID;EDWARDS, RM
使用一种新的方法描述了HIV Tat蛋白基因的合成，该方法利用了合成长度超过100bp的寡核苷酸的能力。它涉及到合成覆盖所需基因的一条完整链的大分子寡聚体，并使用小的互补桥联和接头寡核苷酸来指导大分子寡聚体的组装和克隆。连接到克隆载体后，部分单链中间产物直接转化到受体细菌宿主中，在那里修复质粒。合成的TAT基因已经在HeLa细胞中表达，并被证明能反式激活TAR+，但不能激活TAR-HIV LTR-CAT结构。
The synthesis of a gene for the HIV TAT protein is described using a novel approach that capitalises on the ability to synthesise oligonculeotides 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 intemrediate 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.