高压电场对大肠杆菌的诱变机制研究
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- 英文论文题名：Mutagenic Mechanism Study of High Voltage Electric Field on Escherichia coli
- 论文作者：宋智青 ; 丁昌江 ; 梁运章
- 英文论文作者：SONG Zhiqing ; DING Changjiang ; LIANG Yunzhang ; Physical Experiment Center ; College of Science ; Inner Mongolia University of Technology ; College of Physical Science and Technology ; Inner Mongolia University
- 年：2015
- 作者机构：内蒙古工业大学理学院;内蒙古大学物理科学与技术学院;
- 论文关键词：高压电场 ; 诱变效应 ; 大肠杆菌 ; 分子机制
- 英文论文关键词：High voltage electric field ; Mutagenic effects ; Escherichia coli ; Molecular mechanisms
- 会议召开时间：2015-08-12
- 会议录名称：静电放电：从地面新技术应用到空间卫星安全防护—中国物理学会第二十届全国静电学术会议论文集
- 语种：中文
- 分类号：Q937;O441.4
- 学会代码：WLJD
- 会议名称：静电放电 ; 从地面新技术应用到空间卫星安全防护—中国物理学会第二十届全国静电学术会议
- 会议地点：中国河北石家庄
- 主办单位：中国物理学会静电专业委员会
- 学会名称：中国物理学会静电专业委员会
- 页数：5
- 文件大小：401k
- 原文格式：D
- 会议级别：全国
摘要
为了研究不同高压静电场对大肠杆菌的诱变效应,探讨高压电场诱变效应的可能分子生物学机制和物理学机制,将大肠杆菌K12 W3110野生型菌株干燥菌膜用高压直流、交流、半波整流平行铝板电场,以及针-板高压电晕电场处理,后分别涂布P-gal和LB平皿,考察不同电场的存活率以及诱变率,后筛选高压电晕电场诱变的大肠杆菌突变株,绘制lac I基因突变谱,探讨高压电晕电场诱变的分子生物学机制。结果表明:高压直流、交流、半波整流平板电场处理大肠杆菌K12,3种电场在低剂量处理时(1.5kV/cm时)都表现为对大肠杆菌K12的刺激效应,随着电场剂量增大,3种电场对大肠杆菌K12逐步变为抑制效应。其中直流平板电场的效应最为明显。用高压电晕电场处理大肠杆菌,在电压为6kV,极距为6cm时,突变率为31.2×10~(-6),分别是自发突变(1.2×10~(-6))和对照(2.5×10~(-6))的26倍和12倍。从突变谱看,高压电晕电场组还发现了自发突变组中没有发现的A:T→T:A的颠换和280bp的大片段缺失,同时还有G:C→A:T的增加。分析表明:高压直流、交流、半波整流平板电场对大肠杆菌具有一定的诱变效应,但是其诱变效应不明显。高压电晕电场是一种损伤低、突变率高,突变谱广的很好的诱变剂。从lac I基因的突变谱看,高压电晕电场对大肠杆菌的诱变效应是是由于电场作用引起大肠杆菌SOS反应而导致的;电晕电场和平板电场对大肠杆菌的诱变率不同说明,电晕电场产生的常压低温等离子体对大肠杆菌细胞的损伤,是造成大肠杆菌突变的主要原因。
In order to study of mutagenic effects of several high voltage electric fields,studies of possible mechanisms of high voltage corona electric field(HVCEF).Escherichia coli K12 W3110 was treated 10 min by high voltage DC,AC,half-wave rectification electric fields,The distance between the electrodes is 5cm,and HVCEF,The distance between the electrodes is 6cm.The survival rates and mutation rates was studied.Then,mutant strains of HVPEF treated was selected,after PCR amplification of lac I,mutational spectrum was completed,and we proposed possible mechanisms of HVCEF.The result indicated that the high voltage DC,AC,and half-wave rectification electric fields all induced stimulative effects at low dosage,and with the dosage increasing these three plate electric fields induced inhibitory effects.Among three electric fields,the most evident effects were induced by DC plate electric field.At the field intensity of 4.5 k V/cm,the mutation rate reaches maximum,5.8×10~(-6),which is 2.32 times more than the controls.Escherichia coli K12 W3110 was treated by high voltage corona electrostatic field,The mutant frequency reached31.2×10~(-6) at voltage of 6k V,which was about 26-fold over the background(1.2×10~(-6))and 12-fold over the controls(2.5×10~(-6)).The spectra of our HVCEF exposure group with increasing G:C to A:T transitions and A:T to T:A transversions and large deletion fragment of 280bp.Although these three plate electric field have mutagenic effects on E.coli,but the mutation rates seem not obvious.HVCEF could act as an effective mutagen caused high mutation rate and wide mutational spectrum with low damage.The HVCEF exposure may induce SOS response.The mutation rates were different between plate electric field and corona electric field,it indicated that the atmospheric pressure plasma generated by corona electric field cause the Escherichia coli cells damage,and it was a major cause of Escherichia coli mutant..
In order to study of mutagenic effects of several high voltage electric fields,studies of possible mechanisms of high voltage corona electric field(HVCEF).Escherichia coli K12 W3110 was treated 10 min by high voltage DC,AC,half-wave rectification electric fields,The distance between the electrodes is 5cm,and HVCEF,The distance between the electrodes is 6cm.The survival rates and mutation rates was studied.Then,mutant strains of HVPEF treated was selected,after PCR amplification of lac I,mutational spectrum was completed,and we proposed possible mechanisms of HVCEF.The result indicated that the high voltage DC,AC,and half-wave rectification electric fields all induced stimulative effects at low dosage,and with the dosage increasing these three plate electric fields induced inhibitory effects.Among three electric fields,the most evident effects were induced by DC plate electric field.At the field intensity of 4.5 k V/cm,the mutation rate reaches maximum,5.8×10~(-6),which is 2.32 times more than the controls.Escherichia coli K12 W3110 was treated by high voltage corona electrostatic field,The mutant frequency reached31.2×10~(-6) at voltage of 6k V,which was about 26-fold over the background(1.2×10~(-6))and 12-fold over the controls(2.5×10~(-6)).The spectra of our HVCEF exposure group with increasing G:C to A:T transitions and A:T to T:A transversions and large deletion fragment of 280bp.Although these three plate electric field have mutagenic effects on E.coli,but the mutation rates seem not obvious.HVCEF could act as an effective mutagen caused high mutation rate and wide mutational spectrum with low damage.The HVCEF exposure may induce SOS response.The mutation rates were different between plate electric field and corona electric field,it indicated that the atmospheric pressure plasma generated by corona electric field cause the Escherichia coli cells damage,and it was a major cause of Escherichia coli mutant..
引文
[1]谷卓,那日,石薇,等.高压电晕电场对黄霉素产生菌诱变效应[J].核农学报,2012,26:0740-0745.GU Zhuo,NA Ri,SHI Wei,et al.Effect of corona discharge on flavomycin producting strain[J].Acta Agriculturae Nucleatae Sinica,2012,26:0740-0745.
[2]Cramariuca R.,Donescub V.,Popac M.,et al.The biological effect of the electrical field treatment on the potato seed:agronomic evaluation[J].Journal of Electrostatic,2005,63:837-846.
[3]高雪红,吴俊林.高压静电场在农业中的应用及其作用机理的物理微观解析[J].现代生物医学进展,2008,8:567-570.GAO Xuehong,WU Junlin.Application of High voltage electrostatic field in agriculture and physical microcosmic analysis of its mechanism[J].Progress in modern biomedicine,2008,8:567-570.
[4]Palanimuthu V.,Rajkumar P.,Orsat V.,et al.Improving cranberry shelf-life use high voltage electric field treatment[J].Journal of Food Engineering,2009,90:365-371.
[5]Wartenberg M.,Wirtz N.,Grob A.,et al.Direct current electrical fields induce apoptosis in oral mucosa cancer cells by NADPH oxidase-derived reactive oxygen species[J].Bioelectromagnetics,2008,29:47-54.
[6]耿读艳,徐桂芝,万晓伟,等.模拟输电环境电磁场对动物血清和细胞超微结构的影响[J].高电压技术,2013,39(1):163-174..GENG Duyan,XU Guizhi,WAN Xiaowei,et al.Effect of Electromagnetic Fields in Simulated Transmission Environment on Animal Serum and Cell Ultrastructure[J].High Voltage Engineering,2013,39(1):163-174.
[7]习岗,杨运经,刘锴,等.基于植物电位波动的极低频脉冲电场对绿豆幼苗生长的影响[J].高电压技术2012,38(1):199-204.XI Gang,YANG Yunjing,LIU Kai,et al.Effect of Extremely Low Frequency Pulsed Electric Field Based on Plant Potential Fluctuations on Growth of Mung Bean[J].High Voltage Engineering,2012,38(1):199-204.
[8]习岗,刘锴,杨运经,等.基于玉米幼苗电位波动的脉冲电场生物学效应[J].高电压技术,2013,39(1):129-134.XI Gang,LIU Kai,YANG Yunjing,et al.Biological Effects of Pulsed Electric Field Based on Potential Fluctuations in Maize Seedlings[J].High Voltage Engineering,2012,39(1):129-134.
[9]Mi Y.,Yao C.,Li C.,et al.Caspase-3 Activation by Exponential Decay Nanosecond Pulsed Electric Fields on Tumor-Bearing BALB/c Nude Mice In Vivo[J].IEEETransactions on Plasma Science,2010,38(8):1963-1971.
[10]Pavlin M.,Leben V.,Miklav?i?D.Electroporation in dense cell suspension-Theoretical and experimental analysis of ion diffusion and cell permeabilization[J].Biochimica et Biophysica Acta,2007,1770:12-23.
[11]Miller JH Experiments in Molecular Genetics[M].Cold Spring Harbor Laboratory,New York.1972.
[12]Miller JH.A Short Course in Bacteria Genetics[M].Cold Spring Harbor Laboratory,New York.1992.
[13]Ono T,Negishi K and Hayatsu H.Spectra of superoxide-induced mutations in the lac I gene of a wild-type and a mut M strain of Escherichia coli K-12[J].Mutation Research,1995,326:175-183.
[14]宋智青,罗辽复,梁运章.高压电晕静电场对大肠杆菌K12诱变效应初步研究[J].北京理工大学学报,2009,29(S2):93-95.SONG Zhiqing,LUO Liaofu,LIANG Yunzhang.A pilot study of mutagenic effects of high-voltage prickle electrostatic field on Escherichia coli K12[J].Transactions of Beijing Institute of Technology,2009,29(S2):93-95.
[15]Murata-Kamiya N,Kamiya H,Kaji H,Kasai H.Mutational specificity of glyoxal,a product of DNA oxidation,in the lac Igene of wild type Escherichia coli W3110[J],Mutation Research,1997,377:255-262.
[16]Yoshinori Tanaka,et al.Induction of mutation in Escherichia coli by freeze-drying[J].Apply Environment Microbiology,1979,37(3):369-372.
[17]Shoji Asada,et al.Mutation induced by drying of Escherichia coli on a hydrophobic filter membrane[J].Apply Environment Microbiology,1980,40:274-281.
[18]J.H.Miller,K.B.Low.Specificity of mutagenesis resulting from the induction of the SOS system in the absence of mutagenic treatment[J].Cell,1984,37:675-682.
[19]F.Yatagai,J.Michael,H.Low,B.W.Glickman.Specificity of SOS mutagenesis in native M131ac I phage[J].Journal of Bacteriology,1991,7996-7999.
[20]关志成,杨小卫,王黎明,等.脉冲电磁场对骨质疏松的生物效应[J].高电压技术,2007,33(2):1-6.GUANG Zhi-cheng,YANG Xiao-wei,WANG Li-ming,et al.Bio-effects of pulsed electromagnetic fields on osteoporosis[J].High Voltage Engineering,2007,33(2):1-6.
[21]Nelson D A,Zia S,Whipple R L.Corona discharge effects on heat transfer and pressure drop in tube flows[J].Journal of Enhanced Heat Transfer,2000,7(2):81-95.
[22]Cooke M.S.,Evans M.D.,Dizdaroglu I M.,et al.Oxidative DNAdamage:mechanisms,mutation,and disease[J].Faseb Journal,2003,17:1195-1214.
[23]Gaunt L.F.,Beggs C.B.,Georghiou G.E.Bactericidal action of the reactive species produced by gas-discharge nonthermal plasma at atmospheric pressure:a review[J].IEEE Transactions Plasma Science,2006,34:1257-1269.
[24]May K.,Necdet A.The Effect of Atmospheric Pressure Plasma Corona Discharge on p H,Lipid Content and DNA of Bacterial Cells[J].Plasma Science and Technology,2011,13:99-105.
[2]Cramariuca R.,Donescub V.,Popac M.,et al.The biological effect of the electrical field treatment on the potato seed:agronomic evaluation[J].Journal of Electrostatic,2005,63:837-846.
[3]高雪红,吴俊林.高压静电场在农业中的应用及其作用机理的物理微观解析[J].现代生物医学进展,2008,8:567-570.GAO Xuehong,WU Junlin.Application of High voltage electrostatic field in agriculture and physical microcosmic analysis of its mechanism[J].Progress in modern biomedicine,2008,8:567-570.
[4]Palanimuthu V.,Rajkumar P.,Orsat V.,et al.Improving cranberry shelf-life use high voltage electric field treatment[J].Journal of Food Engineering,2009,90:365-371.
[5]Wartenberg M.,Wirtz N.,Grob A.,et al.Direct current electrical fields induce apoptosis in oral mucosa cancer cells by NADPH oxidase-derived reactive oxygen species[J].Bioelectromagnetics,2008,29:47-54.
[6]耿读艳,徐桂芝,万晓伟,等.模拟输电环境电磁场对动物血清和细胞超微结构的影响[J].高电压技术,2013,39(1):163-174..GENG Duyan,XU Guizhi,WAN Xiaowei,et al.Effect of Electromagnetic Fields in Simulated Transmission Environment on Animal Serum and Cell Ultrastructure[J].High Voltage Engineering,2013,39(1):163-174.
[7]习岗,杨运经,刘锴,等.基于植物电位波动的极低频脉冲电场对绿豆幼苗生长的影响[J].高电压技术2012,38(1):199-204.XI Gang,YANG Yunjing,LIU Kai,et al.Effect of Extremely Low Frequency Pulsed Electric Field Based on Plant Potential Fluctuations on Growth of Mung Bean[J].High Voltage Engineering,2012,38(1):199-204.
[8]习岗,刘锴,杨运经,等.基于玉米幼苗电位波动的脉冲电场生物学效应[J].高电压技术,2013,39(1):129-134.XI Gang,LIU Kai,YANG Yunjing,et al.Biological Effects of Pulsed Electric Field Based on Potential Fluctuations in Maize Seedlings[J].High Voltage Engineering,2012,39(1):129-134.
[9]Mi Y.,Yao C.,Li C.,et al.Caspase-3 Activation by Exponential Decay Nanosecond Pulsed Electric Fields on Tumor-Bearing BALB/c Nude Mice In Vivo[J].IEEETransactions on Plasma Science,2010,38(8):1963-1971.
[10]Pavlin M.,Leben V.,Miklav?i?D.Electroporation in dense cell suspension-Theoretical and experimental analysis of ion diffusion and cell permeabilization[J].Biochimica et Biophysica Acta,2007,1770:12-23.
[11]Miller JH Experiments in Molecular Genetics[M].Cold Spring Harbor Laboratory,New York.1972.
[12]Miller JH.A Short Course in Bacteria Genetics[M].Cold Spring Harbor Laboratory,New York.1992.
[13]Ono T,Negishi K and Hayatsu H.Spectra of superoxide-induced mutations in the lac I gene of a wild-type and a mut M strain of Escherichia coli K-12[J].Mutation Research,1995,326:175-183.
[14]宋智青,罗辽复,梁运章.高压电晕静电场对大肠杆菌K12诱变效应初步研究[J].北京理工大学学报,2009,29(S2):93-95.SONG Zhiqing,LUO Liaofu,LIANG Yunzhang.A pilot study of mutagenic effects of high-voltage prickle electrostatic field on Escherichia coli K12[J].Transactions of Beijing Institute of Technology,2009,29(S2):93-95.
[15]Murata-Kamiya N,Kamiya H,Kaji H,Kasai H.Mutational specificity of glyoxal,a product of DNA oxidation,in the lac Igene of wild type Escherichia coli W3110[J],Mutation Research,1997,377:255-262.
[16]Yoshinori Tanaka,et al.Induction of mutation in Escherichia coli by freeze-drying[J].Apply Environment Microbiology,1979,37(3):369-372.
[17]Shoji Asada,et al.Mutation induced by drying of Escherichia coli on a hydrophobic filter membrane[J].Apply Environment Microbiology,1980,40:274-281.
[18]J.H.Miller,K.B.Low.Specificity of mutagenesis resulting from the induction of the SOS system in the absence of mutagenic treatment[J].Cell,1984,37:675-682.
[19]F.Yatagai,J.Michael,H.Low,B.W.Glickman.Specificity of SOS mutagenesis in native M131ac I phage[J].Journal of Bacteriology,1991,7996-7999.
[20]关志成,杨小卫,王黎明,等.脉冲电磁场对骨质疏松的生物效应[J].高电压技术,2007,33(2):1-6.GUANG Zhi-cheng,YANG Xiao-wei,WANG Li-ming,et al.Bio-effects of pulsed electromagnetic fields on osteoporosis[J].High Voltage Engineering,2007,33(2):1-6.
[21]Nelson D A,Zia S,Whipple R L.Corona discharge effects on heat transfer and pressure drop in tube flows[J].Journal of Enhanced Heat Transfer,2000,7(2):81-95.
[22]Cooke M.S.,Evans M.D.,Dizdaroglu I M.,et al.Oxidative DNAdamage:mechanisms,mutation,and disease[J].Faseb Journal,2003,17:1195-1214.
[23]Gaunt L.F.,Beggs C.B.,Georghiou G.E.Bactericidal action of the reactive species produced by gas-discharge nonthermal plasma at atmospheric pressure:a review[J].IEEE Transactions Plasma Science,2006,34:1257-1269.
[24]May K.,Necdet A.The Effect of Atmospheric Pressure Plasma Corona Discharge on p H,Lipid Content and DNA of Bacterial Cells[J].Plasma Science and Technology,2011,13:99-105.