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原核表达重组质粒pUCS7-CTP-PTEN的构建及鉴定
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原核表达重组质粒pUC57-CTP-PTEN的构建及鉴定
进
喻备，黄媛，胡海峰"，汪自力，杨
（编校：荫明）
Construction and identification of theprokaryotic expression recombinantplasmid pUc57-CTP-PTEN
Yu Bei', Huang Yuan”, Hu Haifeng" , Wang Zili' , Yang Jin'
'Department of Urology; Deparment of Clinical LaboratoryAffliated Hospital of Chengdu Unitersity,Sichuan Chengdu 610081,China
Objective: To construct prokaryotic expression recombinant plasmid pUC57 - CTP - PTEN of CTP
【Abstract】
gene and PTEN gene,and prepare for the follow up study of its function, Methods : Gene CTP PTEN was synthe-sized and cloned into the prokaryotie expression vector pUCS7 to construct the recombinant plasmid pUC57 CTP -PTEN, Then it was transformed into E, coli DH5o. The recombinant plasmid was identified by restriction enzyme di-gestion and DNA sequencing. The expression product was identified by Westerm blotting assay. Results : Gene CTP PTEN was successfully synthesized. The recombinant plasmid pUC57 CTP - PTEN was identified that it was suc-cessfully transformed into E. coli DH5α and expressed by restriction enzyme digestion,DNA sequencing and Westem blotting. Conclusion: The recombinant plasmid pUC57 CTP PTEN was successfully constructed and expressed in E. coli DH5α.
[Key words]plasmid pUC57,CTP,PTEN,construction,identification
Moderm Oncology 2017,25(04) :0528 0530
【摘要】目的：合成基因CTP及PTEN基因，构建其原核表达重组质粒pUC57-CTP-PTEN，为后续研究其功能做准备。方法：合成基因CTP-PTEN，将该基因定向克隆到原核表达载体pUC57中，构建pUC57-CTP-PTEN重组质粒，转化大肠杆菌DH5α菌株，抽提质粒进行双酶切、测序鉴定后，用Westemblotting检验表达。结果：测序鉴定CTP-PTEN基因合成成功。经双酶切、测序鉴定证实重组质粒pUC57-CTP-PTEN成功转人DH5α，Westemblotting检验pUC57-CTP-PTEN成功表达。结论：成功构建了重组质粒pUC57-CTP
-PTEN,并在大肠杆菌DH5α中成功表达。【关键词】质粒pUC57;CTP;PTEN；构建；鉴定
【中图分类号】R730.1
【文献标识码】A
【文章编号】16724992(2017)040528-03
【收稿日期】【修回日期】【基金项目】
【作者单位】【作者简介】【通讯作者】
2016 08 14
DOI:10. 3969/j. issn. 1672 4992. 2017. 04, 06
2016 08 29
成都市医学科研课题（编号：2014001）；成都学院校青年基金项日（编号：2014XJ208）：成都大学附属医院院内课题（编号： 2014016)
"成都大学附属医院泌尿外科；"检验科，四川成都
610081
喻备（1986-），男，四川成都人，医师，硕士，主要从事泌尿系肿瘤生物治疗研究。E-mail：tuy1013@163.com
杨进（1974－），男，四川成都人，博士，副主任医师，副教投，硕士研究生导师，主要从事泌尿系肿瘤研究及泌尿系生物组织工程研究。Email;dr.jinyang@163.com