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大肠杆菌天冬氨酸酶基因(aspA)的克隆

Cloning of the aspartase gene (aspA) of Escherichia coli.

作者信息

Guest J R, Roberts R E, Wilde R J

出版信息

J Gen Microbiol. 1984 May;130(5):1271-8. doi: 10.1099/00221287-130-5-1271.

Abstract

The aspartase gene (aspA) of Escherichia coli has been isolated in two plasmids, pGS73 and pGS94, which contain segments of bacterial DNA (12.5 and 2.8 kb, respectively) inserted into the tet gene of the vector pBR322. The plasmids were constructed by sequential sub-cloning from a larger ColE1-frd+ hybrid plasmid. The location of the aspA gene confirmed predictions based on a correlation between the genetic and restriction maps of the corresponding region. The aspartase activities of plasmid-containing aspA mutants were amplified four- to sixfold relative to aspA+ parental strains. The aspA gene product was tentatively identified as a polypeptide of Mr 55 000, which is somewhat larger than previous estimates (Mr 45000 to 48000) for aspartase.

摘要

大肠杆菌的天冬氨酸酶基因(aspA)已在两种质粒pGS73和pGS94中分离出来,这两种质粒分别含有插入载体pBR322的tet基因中的细菌DNA片段(分别为12.5 kb和2.8 kb)。这些质粒是通过从一个更大的ColE1 - frd + 杂种质粒进行连续亚克隆构建而成的。aspA基因的定位证实了基于相应区域遗传图谱和限制酶切图谱之间相关性所做的预测。相对于aspA + 亲本菌株,含aspA突变体的质粒中天冬氨酸酶活性扩增了4至6倍。aspA基因产物初步鉴定为分子量55000的多肽,这比先前对天冬氨酸酶的估计值(分子量45000至48000)略大。

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