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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2017, Vol. 37 Issue (6): 78-85    DOI: 10.13523/j.cb.20170612
技术与方法     
ARTP辐照对磷脂酶A1重组质粒的诱变效应
杨建伟, 薛正莲, 朱昊, 杨蒙, 王洲
安徽工程大学生物与化学工程学院 微生物发酵安徽省工程技术研究中心 芜湖 241000
Study on the Mutagenic Effect of Phospholipase A1 Recombinant Plasmid by ARTP
YANG Jian-wei, XUE Zheng-lian, ZHU Hao, YANG Meng, WANG Zhou
Institute of Biologic & Chemical Engineering of Anhui Polytechnic University, Microorganism Fermentation Engineering and Technology Research Center of Anhui Province, Wuhu 241000, China
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摘要： 利用常压室温等离子体诱变技术（ARTP）处理磷脂酶A1重组质粒 pET28a-plaB，经琼脂糖凝胶电泳检测发现随着处理时间的延长，质粒超螺旋结构逐渐转变为开环及线性结构。对处理后的质粒转化到BL21宿主菌中，在特定选择培养基中检出突变株转化子，结果表明，质粒处理时间与转化率成反比，与突变率成正比，在60 S达到最优诱变值。挑选圈径比较大的转化子测定其酶活，结果显示突变株（12）最高酶活为12.8 U/ml，与原始菌株（CK）4.8 U/ml相比，提高了2.67倍。对两菌株测序比对，发现碱基突变率为0.74%，且大都集中在A→G和C→T。ARTP对离体质粒具有较好的诱变效应。
关键词： 常压室温等离子体诱变技术;  磷脂酶A1;  诱变;  质粒    
Abstract: The Atmospheric and Room Temperature Plasma (ARTP) was used to treat the recombinant plasmid pET28a-plaB for the expression of phospholipase A1, agarose gel electrophoresis detection showed that with the prolonging of treatment time, the supercoiled plasmid gradually transformed into open loop and linear structure. Then the treatment of plasmid transformed into host strain BL21, the mutant transformants were detected in particular selective medium, the results show that the processing time is directly proportional to the conversion rate and inversely proportional to the rate of mutation, mutation 60s achieved with the optimal value. Selected of the transformants with large circle diameter to detect its enzyme activity, the results showed that the highest enzyme activity of (12) could reach 12.8 U/ml, which increased to 2.67 times than that of the original strain CK(4.8 U/mL). Blasting the sequence of the two strains, the base mutation rate reached 0.74% and the most was concentrated in A→G and C→T. ARTP has a significant mutagenic effect on the in vitro plasmid.
Key words: ARTP    Mutant    Phospholipase A1    Plasmid
收稿日期: 2016-12-02 出版日期: 2017-06-25
ZTFLH:  Q691  
基金资助: 国家自然科学基金（31471615）、安徽省自然科学基金（11040606M81）资助项目
通讯作者: 薛正莲     E-mail: xuezhen0851@sina.com
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引用本文:

杨建伟, 薛正莲, 朱昊, 杨蒙, 王洲. ARTP辐照对磷脂酶A1重组质粒的诱变效应[J]. 中国生物工程杂志, 2017, 37(6): 78-85.

YANG Jian-wei, XUE Zheng-lian, ZHU Hao, YANG Meng, WANG Zhou. Study on the Mutagenic Effect of Phospholipase A1 Recombinant Plasmid by ARTP. China Biotechnology, 2017, 37(6): 78-85.

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https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20170612    或    https://manu60.magtech.com.cn/biotech/CN/Y2017/V37/I6/78

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