Harms N, de Vries G E, Maurer K, Hoogendijk J, Stouthamer A H
J Bacteriol. 1987 Sep;169(9):3969-75. doi: 10.1128/jb.169.9.3969-3975.1987.
A genomic clone bank of Paracoccus denitrificans DNA has been constructed in the expression vector set pEX1, pEX2, and pEX3. Screening of this clone bank with antibodies raised against P. denitrificans methanol dehydrogenase resulted in the isolation of a clone, pNH3, that synthesized methanol dehydrogenase cross-reactive proteins. The nucleotide sequence of the P. denitrificans DNA fragment inserted in this clone has been determined and shown to contain the full methanol dehydrogenase structural gene. DNA cross-hybridization was found with DNA fragments which have been reported to contain the methanol dehydrogenase structural genes from Methylobacterium sp. strain AM1 and Methylobacterium organophilum.
利用表达载体pEX1、pEX2和pEX3构建了反硝化副球菌DNA的基因组克隆文库。用针对反硝化副球菌甲醇脱氢酶产生的抗体筛选该克隆文库,分离出一个克隆pNH3,它能合成与甲醇脱氢酶交叉反应的蛋白质。已确定插入该克隆的反硝化副球菌DNA片段的核苷酸序列,并表明其包含完整的甲醇脱氢酶结构基因。发现与据报道含有甲基杆菌属菌株AM1和嗜有机甲基杆菌甲醇脱氢酶结构基因的DNA片段存在DNA交叉杂交。