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OALib Journal期刊
ISSN: 2333-9721
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大肠杆菌arog基因的定点突变及与trpba基因的串联表达
DOI: 10.3724/SP.J.1145.2013.00817, PP. 817-821
Keywords: 色氨酸,大肠杆菌,arog基因,trpba基因,基因定点突变
Abstract:
在大肠杆菌中arog基因编码的3-脱氧-d-阿拉伯庚酮糖-7-磷酸合成酶（dahps）与trpba基因编码的色氨酸合成酶（tsase）是色氨酸合成的关键酶，为了提高大肠杆菌生产色氨酸的产量，利用soe-pcr方法对arog基因进行632位碱基定点突变，获得抗反馈抑制的定点突变arogfbr基因，并与扩增获得trpba基因串联于petac表达载体上共表达,获得重组菌escherichiacolijm109/petac-arogfbr-tac-trpba，同时构建对照菌株e.colijm109/petac-arog-tac-trpba。两种串联表达质粒重组菌的dahps酶活性，分别比含空载体出发菌株相应酶的活性提高4.3倍和3.8倍，突变dahps酶活提高1.13倍。摇瓶发酵表明，所构建的两菌株与出发菌株色氨酸产量相比较，分别提高8.23倍和11.47倍，带突变arogfbr基因的菌株比对照菌株色氨酸产量提高1.4倍，色氨酸产量明显提高，说明突变菌株能够有效解除苯丙氨酸的反馈抑制作用。图5表2参15
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