A CLONED CYANOBACTERIAL GENE FOR GLUTAMINE-SYNTHETASE FUNCTIONS IN ESCHERICHIA-COLI, BUT THE ENZYME IS NOT ADENYLYLATED
A CLONED CYANOBACTERIAL GENE FOR GLUTAMINE-SYNTHETASE FUNCTIONS IN ESCHERICHIA-COLI, BUT THE ENZYME IS NOT ADENYLYLATED
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DOI：
10.1073/pnas.78.6.3393
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发表时间：
1981-01-01
期刊：
影响因子：
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通讯作者：
HASELKORN, R
中科院分区：
文献类型：
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作者：
FISHER, R;TULI, R;HASELKORN, R
鱼腥藻7120谷氨酰胺合成酶的编码序列[L-谷氨酸：氨连接酶(ADP形成)，EC 6.3.1.2]包含在一个7.5千碱基对(KBP)的HindIII片段中，该片段已通过空斑杂交获得。蓝藻基因的杂交探针是含有E.coliK-12的glnA基因的重组质粒。克隆的鱼腥藻片段含有glnA基因的证据包括与大肠杆菌glnA缺失突变体的互补，以及克隆的鱼腥藻片段所产生的酶与从鱼腥藻7120纯化的谷氨酰胺合成酶的免疫学鉴定。异源双链分析表明，鱼腥藻7120的7.5kbp片段与编码大肠杆菌谷氨酰胺合成酶的11kbp的大肠杆菌片段之间有0.65kbp的同源性。对鱼腥藻glnA基因活性的研究表明，蓝藻基因在大肠杆菌中是不可阻遏的，鱼腥藻7120谷氨酰胺合成酶在大肠杆菌中不是腺基化的。
The coding sequence for Anabaena 7120 glutamine synthetase [L-glutamate:ammonia ligase (ADP-forming), EC 6.3.1.2] are contained within a 7.5-kilobase-pair (kbp) HindIII fragment that has been cloned by plaque hybridization. The hybridization probe for the cyanobacterial gene was a recombinant plasmid containing the glnA gene from E. coli K-12. Evidence that the cloned Anabaena fragment contains the glnA gene includes complementation of a glnA deletion mutant of E. coli and immunological identity of the enzyme produced by the cloned Anabaena fragment in E. coli with glutamine synthetase purified from Anabaena 7120. Heteroduplex analysis reveals 0.65 kbp of homology between the 7.5-kbp Anabaena 7120 fragment and an 11-kbp E. coli fragment that codes for E. coli glutamine synthetase. Studies of Anabaena glnA gene activity in E. coli suggest that the cyanobacterial gene is not repressible and that the Anabaena 7120 glutamine synthetase is not adenylylated in E. coli.