CHARACTERIZATION OF PHOSPHINOTHRICIN ACETYLTRANSFERASE AND C-TERMINAL ENZYMATICALLY ACTIVE FUSION PROTEINS
CHARACTERIZATION OF PHOSPHINOTHRICIN ACETYLTRANSFERASE AND C-TERMINAL ENZYMATICALLY ACTIVE FUSION PROTEINS
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DOI：
10.1016/0378-1119(91)90534-i
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发表时间：
1991-06-15
期刊：
影响因子：
3.5
通讯作者：
LAUWEREYS, M
中科院分区：
文献类型：
--
作者：
BOTTERMAN, J;GOSSELE, V;LAUWEREYS, M
已使用基于翻译偶联的载体系统在大肠杆菌中获得了来自吸水链霉菌的双丙氨膦抗性（bar）的增强表达，所述双丙氨膦抗性赋予对除草剂双丙氨膦和膦丝菌素（PPT）的抗性。将基因产物PPT乙酰转移酶纯化至均一，并分析其酶学性质。具有融合到bar的3 '-末端的基因片段的杂合基因构建体产生具有乙酰转移酶活性的融合蛋白，其对于PPT底物的米氏常数与未修饰的酶相当。bar基因代表特别适合于3 '末端基因融合的可选择和可测定的报告基因。
Enhanced expression of the bialaphos resistance (bar) from Streptomyces hygroscopicus, which confers resistance to the herbicides bialaphos and phosphinothricin (PPT), has been obtained in Escherichia coli using a vector system based on translational coupling. The gene product, PPT acetyltransferase, was purified to homogeneity and its enzymatic properties were analyzed. Hybrid gene constructs with gene fragments fused to the 3'-terminus of bar yield fusion proteins having acetyltransferase activity, with a Michaelis constant for the PPT substrate comparable to the unmodified enzyme. The bar gene represents a selectable and assayable reporter gene especially suitable for 3'-terminal gene fusions.