来自敏捷硫杆菌的甲胺脱氢酶大亚基编码基因的克隆与测序。
Cloning and sequencing of the gene coding for the large subunit of methylamine dehydrogenase from Thiobacillus versutus.
作者信息
Huitema F, van Beeumen J, van Driessche G, Duine J A, Canters G W
机构信息
Gorlaeus Laboratories, Leiden University, The Netherlands.
出版信息
J Bacteriol. 1993 Oct;175(19):6254-9. doi: 10.1128/jb.175.19.6254-6259.1993.
Abstract
The gene that codes for the alpha-subunit of methylamine dehydrogenase from Thiobacillus versutus, madA, was cloned and sequenced. It codes for a protein of 395 amino acids preceded by a leader sequence of 31 amino acids. The derived amino acid sequence was confirmed by partial amino acid sequencing. The start of the mature protein could not be determined by direct sequencing, since the N terminus appeared to be blocked. Instead, it was determined by electrospray mass spectrometry. Confirmation of the results was obtained by sequencing the N terminus after pyroglutamate aminopeptidase digestion. The sequence is homologous to the Paracoccus denitrificans nucleotide sequence. A second open reading frame, called open reading frame 3, is located immediately downstream of madA.
摘要
克隆并测序了来自敏捷硫杆菌的甲胺脱氢酶α亚基编码基因madA。它编码一个由395个氨基酸组成的蛋白质，前面有一个31个氨基酸的前导序列。通过部分氨基酸测序证实了推导的氨基酸序列。由于N端似乎被封闭，无法通过直接测序确定成熟蛋白的起始位置。相反，通过电喷雾质谱法确定了起始位置。在焦谷氨酸氨肽酶消化后对N端进行测序，获得了结果的证实。该序列与反硝化副球菌的核苷酸序列同源。第二个开放阅读框，称为开放阅读框3，位于madA的紧邻下游。
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