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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2017, Vol. 37 Issue (6): 78-85    DOI: 10.13523/j.cb.20170612
    
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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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     
Received: 02 December 2016      Published: 25 June 2017
ZTFLH:  Q691  
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YANG Jian-wei
XUE Zheng-lian
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WANG Zhou
ZHU Hao
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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/10.13523/j.cb.20170612     OR     https://manu60.magtech.com.cn/biotech/Y2017/V37/I6/78

[1] Zhang X, Zhang X F, Li H P, et al. Atmospheric and room temperature plasma (ARTP) as a new powerful mutagenesis tool. Applied Microbiology and Biotechnology, 2014, 98(12):5387-5396.
[2] Wang L Y, Huang Z L, Li G, et al. Novel mutation breeding method for Streptomyces avermitilis using an atmospheric pressure glow discharge plasma. Journal of Applied Microbiology, 2010, 108(3):851-858.
[3] Li H P, Wang Z B, Ge N, et al. Studies on the physical characteristics of the radio-frequency atmospheric-pressure glow discharge plasmas for the genome mutation of Methylosinus trichosporium. IEEE Transactions on Plasma Science, 2012, 40(11):2853-2860.
[4] Zhang X, Zhang C, Zhou Q Q, et al. Quantitative evaluation of DNA damage and mutation rate by atmospheric and room-temperature plasma (ARTP) and conventional mutagenesis. Applied Microbiology and Biotechnology, 2015, 99(13):5639-5646.
[5] Wang L Y, Li G, Zhao H X, et al. A new method for microbial breeding by atmospheric-pressure, non-equilibrium discharge plasma. Journal of Biotechnology, 2008, 136(4):S295-S296.
[6] Wang X, Lu M, Wang S, et al. The atmospheric and room temperature plasma (ARTP) method on the dextranase activity and structure. International Journal of Biological Macro Molecules, 2014,70(8):284-291.
[7] Fang M, Jin L, Zhang C, et al. Rapid mutation of Spirulina platensis by a new mutagenesis system of atmospheric and room temperature plasmas (ARTP) and generation of a mutant library with diverse phenotypes. PLoS One, 2013, 8(10):e77046.
[8] 苏燕南,薛正莲,陈涛,等.粘质沙雷氏菌PL-06磷脂酶A1基因大肠杆菌优化表达.中国生物工程杂志,2013,33(7):36-42. Su Y N, Xue Z L, Chen T, et al.The optimized phospholipase A1 gene expression of Serratia marcescens PL-06 in E.coli.China Biotechnology,2013,33(7):36-42.
[9] Alaoui M E, Soulère L,Noiriel A, et al.A continuous spectrophotometric assay that distinguishes between phospholipase A1 and A2 activities.Journal of Lipid Research,2016, 57(10):1589-1597.
[10] Ranjan Moharana T. Selective enrichment of omega-3 fatty acids in oils by phospholipase A1.PLoS One,2016,11(3):e0151370.
[11] Bakholdina S I, Tischenko N M, Sidorin E V, et al.Recombinant phospholipase A1 of the outer membrane of Psychrotrophic yersinia pseudotuberculosis:expression, purification, and characterization.Biochemistry, 2016,81(1):47-57.
[12] 吴桂梅. 柱式法和磁珠法提取大肠杆菌中质粒.生物技术世界, 2015,26(11):26-27 Wu G M.Pillar method and magnetic beads method to extract plasmid from E. coli. Biotech World, 2015,26(11):26-27
[13] 刘成君,黄庆,赵莲. 大肠杆菌-枯草杆菌穿梭质粒载体pSUGV4的构建. 四川大学学报(自然科学版),2001,38(2):243-246. Liu C J, Huang Q,Zhao L.Construction of a Escherichia coli-Bacillus subtilis shuttle vector pSUGV4. Journal of Sichuan University (Natural Science Edition),2001,38(2):243-246.
[14] 谢晖.紫外照射对大肠埃希氏菌质粒DNA影响的研究.青海民族大学学报(教育科学版), 2010, 30(5):48-51. Xie H.The influence on Escherichia coli plasmid DNA which caused by SOS repaired system combined with mltraviolet irradiation mutation.Journal of Qinghai Junior Teachers College,2010, 30(5):48-51.
[15] 李莉,杨剑波,李俊,等.离子束辐照对pUC18质粒DNA结构和功能影响的初步研究. 安徽农业科学,1994,04:300-304. Li L, Yang J H, Li J, et al.Research on damage of ion beam irradiated pUC18 plasmid DNA.Journal of AnHui Agricultural Sciences,1994,04:300-304.
[16] 陈璇,刘松,冯岳,等. 常压室温等离子诱变选育L-天冬酰胺酶高产重组菌. 食品与生物技术学报,2016,35(5):485-491. Chen X, Liu S, Feng Y, et al.Screening of high L-asparaginase activity mutants of recombinant bacteria.Journal of Food Science and Biotechnology,2016,35(5):485-491.
[17] 周国雁,郭凤根,张应华,等.一种经济高效的用Silica提取纯化质粒DNA的方法. 生物技术通讯, 2007, 18(5):800-802. Zhou G Y, Guo F G,Zhang Y H,et al.A rapid and high efficiency method for plasmid DNA extraction and purification using silica.Letters in Biotechnology,2007, 18(5):800-802.
[18] Li X, Jin H, Wu Z, et al. An automated process to extract plasmid DNA by alkaline lysis. Applied Microbiology and Biotechnology, 2007, 75(5):1217-1223.
[19] Li G, Li H P, Wang L Y, et al. Genetic effects of radio frequency,atmospheric-pressure glow discharges with helium. Applied Physics Letters, 2008, 92(22):221504.
[20] 何建华, 蒋玮, 吕贝贝, 等. ARTP诱变筛选草菇优良菌株及RAPD分析. 核农学报, 2014, 28(11):1950-1955.
[21] Dong X Y, Xiu Z L, Hou Y M, et al. Enhanced production of 1, 3-propanediol in Klebsiella pneumoniae induced by dielectric barrier discharge plasma in atmospheric air. IEEE Transactions on Plasma Science, 2009, 37(6):920-926.
[22] Lu Y, Wang L, Ma K, et al. Characteristics of hydrogen production of an Enterobacter aerogenes mutant generated by a new atmospheric and room temperature plasma (ARTP). Biochemical Engineering Journal, 2011, 55(1):17-22.
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