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慢生根瘤菌腺苷酸环化酶基因(cya)在大肠杆菌中的分离与表达

Isolation and expression of the Bradyrhizobium japonicum adenylate cyclase gene (cya) in Escherichia coli.

作者信息

Guerinot M L, Chelm B K

出版信息

J Bacteriol. 1984 Sep;159(3):1068-71. doi: 10.1128/jb.159.3.1068-1071.1984.

Abstract

A 5.0-kilobase-pair HindIII fragment of Bradyrhizobium japonicum DNA containing the cya gene which encodes adenylate cyclase was isolated as an insert in pBR322, using marker rescue of the maltose-negative phenotype of an Escherichia coli cya mutant for identification. The isolated B. japonicum DNA fragment was capable of reversing the pleiotropic phenotype of cya mutations when inserted in either orientation in the HindIII site of pBR322. The complemented E. coli strains produced high levels of cyclic AMP. No sequence homology between the B. japonicum cya gene and that of E. coli was detected by hybridization analysis.

摘要

利用大肠杆菌cya突变体麦芽糖阴性表型的标记拯救进行鉴定,从日本慢生根瘤菌DNA中分离出一个包含编码腺苷酸环化酶的cya基因的5.0千碱基对HindIII片段,该片段作为插入片段插入到pBR322中。当以任何一种方向插入到pBR322的HindIII位点时,分离出的日本慢生根瘤菌DNA片段都能够逆转cya突变的多效表型。互补的大肠杆菌菌株产生高水平的环磷酸腺苷。通过杂交分析未检测到日本慢生根瘤菌cya基因与大肠杆菌cya基因之间的序列同源性。

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