Vettakkorumakankav N N, Stevenson K J
Department of Biological Sciences, University of Calgary, Alta., Canada.
Biochem Cell Biol. 1992 Aug;70(8):656-63. doi: 10.1139/o92-101.
We used the N-terminal amino acid sequence of dihydrolipoamide dehydrogenase from Haloferax volcanii, to design and synthesize two oligonucleotide probes that were used to identify and clone a 4.3 kilobase pair (kbp) fragment from MboI restriction endonuclease digestion of Hf. volcanii genomic DNA. The nucleotide sequence of a 1.5-kbp region of this clone was determined and this revealed an open reading frame that translated into a protein with good homology to dihydrolipoamide dehydrogenase from other sources. The first 48 amino acids were identical with the N-terminal sequence data obtained from the purified protein. The complete primary structure of the halophilic dihydrolipoamide dehydrogenase was analyzed in terms of its homologies to dihydrolipoamide dehydrogenases from other sources and its molecular adaptations to high intracellular ionic strength.
我们利用来自嗜盐嗜热栖热菌的二氢硫辛酰胺脱氢酶的N端氨基酸序列，设计并合成了两个寡核苷酸探针，用于从嗜盐嗜热栖热菌基因组DNA的MboI限制性内切酶消化产物中鉴定并克隆一个4.3千碱基对（kbp）的片段。测定了该克隆1.5-kbp区域的核苷酸序列，并发现了一个开放阅读框，其编码的蛋白质与其他来源的二氢硫辛酰胺脱氢酶具有高度同源性。前48个氨基酸与从纯化蛋白获得的N端序列数据一致。从嗜盐二氢硫辛酰胺脱氢酶与其他来源的二氢硫辛酰胺脱氢酶的同源性以及其对高细胞内离子强度的分子适应性方面，分析了其完整的一级结构。
