| N+离子注入诱变筛选胸腺嘧啶缺陷型大肠杆菌 |
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| 引用本文： | 昌国强,欧伶,丁庆豹,郑之明,王丽,袁成凌.N+离子注入诱变筛选胸腺嘧啶缺陷型大肠杆菌[J].化学与生物工程,2011,28(4):49-52,72. |
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| 作者姓名： | 昌国强 欧伶 丁庆豹 郑之明 王丽 袁成凌 |
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| 作者单位： | 1. 华东理工大学生物反应器工程国家重点实验室,上海,200237
2. 中国科学院等离子体物理研究所离子束生物工程学重点实验室,安徽合肥,230031 |
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| 基金项目： | 中国科学院等离子体物理研究所暨安徽省离子束生物工程学重文实验室开放基金资助项目 |
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| 摘 要： | 利用N+离子注入诱变大肠杆菌,经甲氧苄啶的选择性筛选,得到胸腺嘧啶营养缺陷型菌株.通过重组DNA技术将该突变株胸苷酸合成酶(TS)基因连接到载体pMD-18T质粒上,经测序发现TS基因第173个碱基由AT→CG,其相应的氨基酸由谷氨酸(E58)突变为丙氨酸(A58).该突变株的获得为后续研究嘧啶核苷酸的代谢奠定了基础.
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| 关 键 词： | 低能N+离子注入 甲氧苄啶 胸苷酸合成酶 |
Breeding of Thymineless Escherichia Coli Mutated with N+ Ions Implantation |
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| CHANG Guo-qiang,OU Ling,DING Qing-bao,ZHENG Zhi-ming,WANG Li,YUAN Cheng-ling.Breeding of Thymineless Escherichia Coli Mutated with N+ Ions Implantation[J].Chemistry & Bioengineering,2011,28(4):49-52,72. |
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| Authors: | CHANG Guo-qiang OU Ling DING Qing-bao ZHENG Zhi-ming WANG Li YUAN Cheng-ling |
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| Affiliation: | 1.State Key Laboratory of Bioreactor Engineering,East China University of Science and Technology,Shanghai 200237,China;2.Key Laboratory of Ion Bean Bioengineering,Institute of Plasma Physics,Chinese Academy of Sciences,Hefei 230031,China) |
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| Abstract: | Escherichia coli was mutated with N+ions implantation and the thymineless mutants was screened selectively by trimethoprim.The gene encoding thymidylate synthase(TS) from mutant was cloned into vector pMD-18T by recombinant DNA technology.It was found that the 173th base pairs were changed from AT to CG by gene-sequencing,thus the corresponding amino acid was changed from glutamic acid(E58) to alanine(A58).These studies are to provide a good basis for the further study of pyrimidine nucleotide metabolism. |
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| Keywords: | N+ions implantation trimethoprim thymidylate synthase |
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