Altered Cro repressors from engineered mutagenesis of a synthetic cro gene.
Altered Cro repressors from engineered mutagenesis of a synthetic cro gene.
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通过合成 cro 基因的工程诱变改变了 Cro 阻遏蛋白。
DOI：
10.1073/pnas.82.4.1084
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发表时间：
1985
影响因子：
11.1
通讯作者：
Caruthers,MH
中科院分区：
文献类型：
--
作者：
Eisenbeis,SJ;Nasoff,MS;Noble,SA;Bracco,LP;Dodds,DR;Caruthers,MH
编码λ噬菌体的Cro阻遏蛋白的基因的一部分已被化学合成，掺入产生限制性内切酶位点而不改变氨基酸编码序列的碱基对变化。这些限制性内切酶位点用于去除合成cro基因的小片段，并且用携带所需突变的双链体替换所述片段。然后测定以这种方式构建的突变体产生的改变的Cro蛋白在体内与λ操纵基因OR 3的结合。定向进入编码α-3螺旋的cro基因区域的突变产生了改变的Cro蛋白，其对操纵基因DNA具有一系列亲和力。这些变化表明哪些氨基酸在Cro-OR 3复合物形成中起重要作用。
A portion of the gene coding for the Cro repressor protein of bacteriophage lambda has been chemically synthesized, incorporating base pair changes that generate restriction endonuclease sites without altering the amino acid coding sequence. These restriction endonuclease sites were used to remove small segments of the synthetic cro gene and the segments were replaced with duplexes carrying desired mutations. Altered Cro proteins produced by mutants constructed in this manner were then assayed for binding to lambda operator OR3 in vivo. Mutations directed into the region of the cro gene encoding the alpha-3 helix produced altered Cro proteins with a range of affinities for operator DNA. These changes suggest which amino acids play an important role in Cro-OR3 complex formation.