甲基四氢叶酸还原酶_

人类5,10亚甲基四氢叶酸还原酶N端部分的克隆和表达

http://www.100md.com 《第四军医大学学报》 2003年第7期

甲基四氢叶酸还原酶,,甲基四氢叶酸还原酶；克隆；原核表达,0引言,1材料和方法,2结果,3讨论,【参考文献】

     (第四军医大学西京医院：神经内科，检验科，陕西西安 710033)

    Cloning and expression of N terminal human 5, 10methylenetetrahydrofolate reductase code gene

    WAN Qi, SUN WenPing, SU MingQuan, YU YingXin, LI Li, ZANG RuiGuo, GUO YiChuang

    Department of Neurology, Department of Clinical Laboratory, Xijing Hospital, Fourth Military Medical University, Xian 710033, China

    【Abstract】 AIM: To clone human N terminal human 5, 10methylenetetrahydrofolate reductase [MTHFR(N)] code gene, construct the recombinant vector and express its product. METHODS: Human MTHFR(N) code gene was amplified by RTPCR from fetal liver tissue and cloned into vector pGEMT easy, which was sequenced and subcloned into prokaryotic expressive vector pGEX4T2. After a 4hour induction by IPTG, the recombinant Mr 66ku fusion protein GSTMTHFR(N) was expressed and confirmed by SDSPAGE. RESULTS: Human MTHFR(N) gene was cloned. Recombinant expression plasmid pGEX 4T2/ MTHFR(N) was constructed. The expressed fusion protein was about 31% of the total bacterial protein by gel thinlayer chromatographyscanning. CONCLUSION: The human MTHFR(N) gene and prokaryotic expression products have been obtained, which is of great importance for further study on the function of human MTHFR. ......

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