Cloning, nucleotide sequence, and overexpression of the bacteriophage T4 regA gene.
Cloning, nucleotide sequence, and overexpression of the bacteriophage T4 regA gene.
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噬菌体 T4 regA 基因的克隆、核苷酸序列和过表达。
DOI：
10.1073/pnas.82.7.1901
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发表时间：
1985
影响因子：
11.1
通讯作者：
Spicer,EK
中科院分区：
文献类型：
--
作者：
Adari,HY;Rose,K;Williams,KR;Konigsberg,WH;Lin,TC;Spicer,EK
噬菌体T4 regA基因编码一种调控蛋白，该蛋白控制许多T4早期基因的表达，显然是在翻译水平上。已将含有regA结构基因的限制性片段克隆到噬菌体M13中，并测定了核苷酸序列。DNA序列的翻译预测regA蛋白含有122个氨基酸，Mr为14，620。将含有regA基因85%编码序列的DNA片段克隆到噬菌体λ基因启动子PL表达载体pAS 1中，经萘啶酸诱导，获得了高水平的regA截短蛋白。截短的regA蛋白的蛋白质化学研究给出了与regA基因的核苷酸序列一致的结果。随后，将完整的regA基因克隆到质粒pAS 1中并过表达。当T4 45和44蛋白的相应基因与regA基因携带在同一质粒上时，以这种方式产生的regA蛋白调节T4 45和44蛋白的水平。
The bacteriophage T4 regA gene codes for a regulatory protein that controls the expression of a number of T4 early genes, apparently at the level of translation. Restriction fragments containing the regA structural gene have been cloned into phage M13, and the nucleotide sequence has been determined. Translation of the DNA sequence predicted that regA protein contains 122 amino acids, with a Mr of 14,620. A DNA fragment carrying 85% of the coding sequence of regA has been cloned into the phage lambda leftward promoter PL expression vector pAS1, and a high level of truncated regA protein was produced by nalidixic acid induction. Protein chemical studies of the truncated regA protein gave results consistent with the nucleotide sequence of the regA gene. Subsequently, an intact regA gene was cloned into plasmid pAS1 and overexpressed. The regA protein produced in this way regulates the level of T4 45 and 44 proteins when their corresponding genes are carried on the same plasmid as the regA gene.