Wang M D, Buckley L, Berg C M
Department of Molecular and Cell Biology, University of Connecticut, Storrs 06268.
J Bacteriol. 1987 Dec;169(12):5610-4. doi: 10.1128/jb.169.12.5610-5614.1987.
To facilitate molecular analyses of a previously uncharacterized gene involved in alanine synthesis, attempts were made to clone the wild-type allele of this gene, alaA, with a mini-Mu plasmid element used for in vivo cloning. Seventy-six independent Ala+ plasmids were isolated and characterized. Physiological, enzymological, and restriction endonuclease analyses indicated that three different genes, none of them alaA, were cloned. These genes were avtA+, which encodes the alanine-valine transaminase (transaminase C); tyrB+, which encodes the tyrosine-repressible transaminase (transaminase D); and a previously undescribed gene, called alaB, which encodes an alanine-glutamate transaminase.
为便于对参与丙氨酸合成的一个此前未被鉴定的基因进行分子分析,研究人员尝试用一种用于体内克隆的微型Mu质粒元件来克隆该基因(alaA)的野生型等位基因。分离并鉴定了76个独立的Ala⁺质粒。生理、酶学和限制性内切酶分析表明,克隆到的是三个不同的基因,均不是alaA。这些基因分别是:avtA⁺,编码丙氨酸-缬氨酸转氨酶(转氨酶C);tyrB⁺,编码酪氨酸可阻遏的转氨酶(转氨酶D);以及一个此前未描述的基因alaB,编码丙氨酸-谷氨酸转氨酶。