Yamamoto T, Tomiyama M, Mita H, Sode K, Karube I
Research Department, NOK Corporation, Fujisawa, Japan.
FEMS Microbiol Lett. 1990 Jan 1;54(1-3):187-92. doi: 10.1016/0378-1097(90)90280-4.
The hydB gene of Escherichia coli, which is related with the expression of hydrogenase activity, was cloned into the plasmid (pES1). Using the maxicell protein-labeling method, the molecular weight of hydB gene product was estimated. Comparing between the gene products from the mutant strains and that of the hydB genes cloned strains, the molecular weight of the gene product was 35,000 Mr. Similarly, the molecular weight of the gene product of hydA, which had been previously cloned, was determined by maxicell analysis. The molecular weight of hydA gene product was estimated to be 80,000 Mr. Using deletion analysis and Tn1000 insertional inactivation of hydA's function, the hydA coding region was estimated between 2.2 kb and 2.8 kb in a 3.1 kb EcoRI-MluI fragment on the recombinant plasmid pEH3.
将与氢化酶活性表达相关的大肠杆菌hydB基因克隆到质粒(pES1)中。采用最大细胞蛋白标记法对hydB基因产物的分子量进行了估算。通过比较突变菌株和hydB基因克隆菌株的基因产物,该基因产物的分子量为35,000道尔顿。同样,通过最大细胞分析确定了先前克隆的hydA基因产物的分子量,hydA基因产物的分子量估计为80,000道尔顿。利用缺失分析和hydA功能的Tn1000插入失活,估计重组质粒pEH3上3.1 kb EcoRI - MluI片段中hydA编码区在2.2 kb至2.8 kb之间。