酿酒酵母SAM合成酶基因的克隆及表达.pdf

5恤ChineseScience of Nov．2006 TechnologyEducationSymposiumPharmaceutical Engineering 酿酒酵母SAM合成酶基因的克隆及表达 安胜欣王雅琴幸任远 北京化工大学生命科学与技术学院，北京 100029 ·通讯作者：E—mail：yaqinw@mail．buct．edu．cnI 北京市北三环东路15号北京化工大学53号信箱 关键词酿酒酵母；S一腺苷甲硫氨酸合成酶；基因表达 摘要目的：酿酒酵母S一腺苷甲硫氨酸(SAM)合成酶基因在大肠杆菌中的克隆和表达。方法：以酿酒酵母2B一4l的 表达产物主要以包涵体的形式存在。结论：酿酒酵母S一腺苷甲硫氨酸(SAM)合成酶基因可在大肠杆菌中的克隆和表达。 and of expression Cloning S—adenosylmethioninesynthetase f．rom cerevisiae gene Saccharomyces AN xinRENYuanWANG Sheng Yaqin4 ofLifeScienceand ofChemical College Technology,BeijingUniversity Technology，Beijing，100029 words Key Saccharomycescerevisiae；S-adenosylmethioninesynthetase；geneexpression AbstractAhn．*ToCloneand SAM from cerevisiaeinE．coli．Methods：The expresssynthetasegene Saccharomyces geneencoding SAM2was from cerevisiae2B-41chromosomalDNAPCR onthebaseofthe amplifiedSaccharomyces using technique specific from cerevisiaeinGenBank．ThePCR was DNA wasthen primersSaccharomyces productAnalyszedby sequence．Thetarget insertedintothe vector and inE．coli recombinantwas as expressionpET-28aexpressedBL21(DE3)．Result：Theproteinexpressed aninclusionand 23．3％ofthebacterialtotal showedthattherelativemolecular bodyaccumulated叩to protein．SDS·PAGEanalysis wasabout from massoftherecombinant 47KD．ThattosizeofSAM protein equal synthetase．Conclusion：SAMsynthetasegene cerevisiaeWas i