CLONING AND CHARACTERIZATION OF THE HUMAN 5,10-METHENYLTETRAHYDROFOLATE SYNTHETASE-ENCODING CDNA
CLONING AND CHARACTERIZATION OF THE HUMAN 5,10-METHENYLTETRAHYDROFOLATE SYNTHETASE-ENCODING CDNA
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DOI：
10.1016/0378-1119(95)00321-v
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发表时间：
1995-11-20
期刊：
影响因子：
3.5
通讯作者：
JOLIVET, J
中科院分区：
文献类型：
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作者：
DAYAN, A;BERTRAND, R;JOLIVET, J
次甲基四氢叶酸合成酶 (MTHFS) 催化细胞内将 5-甲酰四氢叶酸转化为其他还原叶酸的必要初始代谢步骤。我们分离并测序了人类 MTHFS cDNA，该 cDNA 长 872 bp，编码 23229 Da 的 203 个氨基酸蛋白质。用含有编码 MTHFS 的开放阅读框的 pET11c 质粒转染的大肠杆菌 BL21(DE3)，在 IPTG 诱导后，细菌提取物中的 MTHFS 活性增加了 100 倍。人体组织的 Northern 印迹研究确定 MTHFS mRNA 优先在肝脏中表达，而人类基因组 DNA 的 Southern 印迹分析表明存在单拷贝基因。
Methenyltetrahydrofolate synthetase (MTHFS) catalyses the obligatory initial metabolic step in the intracellular conversion of 5-formyltetrahydrofolate to other reduced folates, We have isolated and sequenced a human MTHFS cDNA which is 872-bp long and codes for a 203-amino-acid protein of 23229 Da. Escherichia coli BL21(DE3), transfected with pET11c plasmids containing an open reading frame encoding MTHFS, showed a 100-fold increase in MTHFS activity in bacterial extracts after IPTG induction. Northern blot studies of human tissues determined that the MTHFS mRNA was expressed preferentially in the liver and Southern blot analysis of human genomic DNA suggested the presence of a single-copy gene.