Heilmann H J, Mägert H J, Gassen H G
Institut für Organische Chemie und Biochemie, Technische Hochschule Darmstadt, Federal Republic of Germany.
Eur J Biochem. 1988 Jun 15;174(3):485-90. doi: 10.1111/j.1432-1033.1988.tb14124.x.
A gene encoding glucose dehydrogenase of Bacillus megaterium M1286 was isolated from a lambda-EMBL3 phage library. It is transcribed and translated in cells of the heterologous organism Escherichia coli by own control regions. The gene is located on a 1126-bp HindIII fragment. Its nucleotide sequence contains 220 bp in the 5' non-coding region, 783 bp in the coding region and 123 bp in the 3' non-coding region. The amino acid sequence, as deduced from the coding region, consists of 261 amino acids and is different from the known protein sequence of glucose dehydrogenase from B. megaterium M1286. [Jany, K. D., Ulmer, W., Fröschle, M. & Pfleiderer, G. (1984) FEBS Lett. 165, 6-10]. By using this gene as a hybridization probe a second glucose dehydrogenase gene was isolated, which was also directly expressed in E. coli. Additionally a DNA region with extended sequence homology to the hybridization probe was identified. This work indicates the existence of at least two independent glucose dehydrogenase genes in B. megaterium M1286. Homologies in the primary structures of the two different glucose dehydrogenases of B. megaterium M1286 and of the corresponding Bacillus subtilis enzyme are discussed.
从λ-EMBL3噬菌体文库中分离出编码巨大芽孢杆菌M1286葡萄糖脱氢酶的基因。它通过自身的调控区在异源生物大肠杆菌细胞中进行转录和翻译。该基因位于一个1126 bp的HindIII片段上。其核苷酸序列在5'非编码区含有220 bp，在编码区含有783 bp，在3'非编码区含有123 bp。从编码区推导的氨基酸序列由261个氨基酸组成，与巨大芽孢杆菌M1286已知的葡萄糖脱氢酶蛋白质序列不同。[扬尼，K.D.，乌尔默，W.，弗罗施勒，M. & 普弗莱德雷尔，G.（1984年）《欧洲生物化学学会联合会快报》165，6 - 10]。通过使用该基因作为杂交探针，分离出了第二个葡萄糖脱氢酶基因，它也能在大肠杆菌中直接表达。此外，还鉴定出了一个与杂交探针具有延伸序列同源性的DNA区域。这项工作表明巨大芽孢杆菌M1286中至少存在两个独立的葡萄糖脱氢酶基因。讨论了巨大芽孢杆菌M1286两种不同葡萄糖脱氢酶以及相应枯草芽孢杆菌酶一级结构中的同源性。
