| 甲基营养菌Methylobacterium sp. MB200中部分一碳代谢相关基因的定位及mtdA和mtdB 基因的克隆研究 |
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| 引用本文： | 宋修鹏,武波,申佩弘,蒋承建,田丹丹,唐咸来. 甲基营养菌Methylobacterium sp. MB200中部分一碳代谢相关基因的定位及mtdA和mtdB 基因的克隆研究[J]. 中国生物工程杂志, 2011, 31(2): 50-55 |
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| 作者姓名： | 宋修鹏 武波 申佩弘 蒋承建 田丹丹 唐咸来 |
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| 作者单位： | 1. 广西大学生命科学与技术学院 南宁 530005;2. 微生物及植物遗传工程教育部重点实验室 南宁 530005;3. 广西亚热带生物资源保护利用重点实验室 南宁 530005;4. 广西壮族自治区科学技术厅 南宁 530005 |
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| 摘 要： | 以甲基营养细菌Methylobacterium sp. MB200为出发菌株,首先利用质粒转座子(pTnMod-RKm’)构建了目的菌株MB200的部分突变体库,获得能够在双抗性MMII板上生长的突变体共11552株,然后在双抗性MMI板上复筛获得不能够利用甲醇的突变体333株(初步认为是一碳代谢途径被破坏的突变株),为一碳代谢相关基因的克隆研究奠定了基础。随后利用TAIL-PCR技术快速准确地从突变体库中定位了部分与一碳代谢相关的基因,并根据pTnMod-RKm’具有复制起始位点能够快速克隆插入位点侧翼序列的特性克隆到mtdA和mtdB基因的全序列并进行了初步分析。
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| 关 键 词： | 甲基营养菌 突变体库 pTnMod-RKm’ TAIL-PCR mtdA mtdB |
| 收稿时间： | 2010-10-13 |
| 修稿时间： | 2010-11-30 |
Location of Some Genes Related to One Carbon Metabolism in Methylobacterium sp.MB200 and Cloning of mtdA and mtdB Genes |
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| SONG Xiu-peng,WU Bo,SHEN Pei-hong,JIANG Cheng-jian,TIAN Dan-dan,TANG Xian-lai. Location of Some Genes Related to One Carbon Metabolism in Methylobacterium sp.MB200 and Cloning of mtdA and mtdB Genes[J]. China Biotechnology, 2011, 31(2): 50-55 |
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| Authors: | SONG Xiu-peng WU Bo SHEN Pei-hong JIANG Cheng-jian TIAN Dan-dan TANG Xian-lai |
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| Affiliation: | SONG Xiu-peng1,2,3 WU Bo1,3 SHEN Pei-hong1,3 JIANG Cheng-jian1,3 TIAN Dan-dan1,3 TANG Xian-lai1,4(1 The College of Iife Science and Technology,Guangxi University,Nanning 530005,China) (2 Key laboratory of Microbial and Plant Genetic Engineering of Ministry of Education,China) (3 Key Laboratory of Subtropical Bioresource Conservation and Utilization of Guangxi,China) (4 The Science and Technology Department of Guangxi,China) |
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| Abstract: | The partial mutants library of Methylobacterium sp.MB200 was constructed by using plasmid pTnMod-RKm' and 11554 mutants were obtained which could survive in MMII containing two antibiotics(Nm and Km).Rescreening results showed that 333 strains could not use methanol as the sole carbon source,these strains were preliminarily considered that their one carbon metabolism way had been destroyed.This laid the foundation for cloning genes related to one carbon metabolism.The location analysis was carried out by TA... |
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| Keywords: | Methylobacterium Mutants library pTnMod-RKm' TAIL-PCR mtdA mtdB |
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