谷氨酸棒杆菌谷氨酰胺合成酶基因的克隆和表达研究.pdf

5mChineseScience ofPharmaceuticalNov．2006 TechnologyEducationSymposium Engineering 谷氨酸棒杆菌谷氨酰胺合成酶 基因的克隆和表达 张春辉”苗明峰2’张少平1’杜娟1’穆瑞瑞’’吴健’)’李季伦” 1)郑州大学生物工程系中国郑州4500012)河南省实验中学中国郑州450002 关键词 谷氯酰胺合成酶；工程菌；基因表达；谷氨酰胺 922 中图分类号TQ 文献标识码A 摘 用IPTG诱导表达后SDS分析，所表达的谷氨酰胺合成酶(glutamine pUCl8-glnA和基因工程菌构建成功。 4 anand oI frIrOmm CloningExllzxpresslonglnAglnAgene 0ryneoac gtutamtcum ZHANGChunhuil’MIA0 Juanl’MURuiruil’WUJianl’LIJilunl’ Yuefeng卸ZHANGShaopin91’DU 1．DepartmentofBioengeering,ZhengzhouUniversity,Zhengzhou450001； 2．HenanProvince School 450002 Experiment Zhengzhou High words Key glutaminesynthetase(GS)；engineeredbacteria；geneexpression；glutamine Abstract the andfunctionof from in Aim：Studyexpression glnAgene Corynebacteriumglutamicum wasusedtO whichwasrecombinedwith 8tOformthevector synthetase pUC- technique amplifyglnAgeneencodingglutamine pUCI DNA vectorWastransformedinto E．coliDH5a induced glnA．Aftersequenceanalysis，the competent strain．Results：Expressionby IPTGWas andtheinducedWas to about40％ofthetotal SDS—PAGE performed proteinprovedoccupy proteinsby analysis．The recombinantbacteriahad incrudeextract4times thanthatofthe DH5astrain．Conehmion：Itis enzymeactivity higher original thattherecombinantvector and E．coliDHSastrainwereconstructed proved expressionpUCl8-glnAengineenng successfully． 谷氨酰胺合成酶在生物体氮代谢中处于关