来自柱状梭菌的N10-甲酰四氢叶酸合成酶基因的序列与表达
Sequence and expression of the gene for N10-formyltetrahydrofolate synthetase from Clostridium cylindrosporum.
作者信息
Rankin C A, Haslam G C, Himes R H
机构信息
Department of Biochemistry, University of Kansas, Lawrence 66045-2106.
出版信息
Protein Sci. 1993 Feb;2(2):197-205. doi: 10.1002/pro.5560020208.
Abstract
Sau3 A and Hind III restriction fragments of Clostridium cylindrosporum genomic DNA were used to isolate clones containing 80% of the N10-H4folate synthetase gene in a 5' fragment and the remaining 20% of the gene in the 3' fragment. These fragments were joined at a common SnaB I restriction site and expressed in Escherichia coli at a level equivalent to what is normally found in C. cylindrosporum. Sequence comparisons show a large degree of homology with genes from two other clostridial species, including a thermophile. Certain conserved sequences found in the three clostridial proteins and in the N10-H4folate synthetase portion of eukaryotic C1-H4folate synthases may represent consensus sequences for nucleotide and H4folate binding.
摘要
圆柱形梭菌基因组DNA的Sau3A和Hind III限制性片段用于分离克隆，这些克隆在5'片段中包含80%的N10-H4叶酸合成酶基因，在3'片段中包含该基因的其余20%。这些片段在一个共同的SnaB I限制性位点处连接，并在大肠杆菌中表达，表达水平与圆柱形梭菌中通常发现的水平相当。序列比较显示与另外两种梭菌属物种（包括一种嗜热菌）的基因有很大程度的同源性。在这三种梭菌蛋白以及真核生物C1-H4叶酸合成酶的N10-H4叶酸合成酶部分中发现的某些保守序列可能代表核苷酸和H4叶酸结合的共有序列。
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