离子液体[BMIm][PF6]增强抗性质粒RP4介导的接合转移基因mRNA的表达水平An Ionic Liquid [BMIm][PF6] Enhanced the mRNA Expression Levels of Conjugative Transfer Genes Mediated by Plasmid RP4
汪庆;罗义;崔玉晓;齐丽英;周烁磊;杨光;
摘要(Abstract):
环境中广泛存在的抗生素和抗生素抗性基因会导致很严重的人类健康风险。在我们前期研究中发现,离子液体1-丁基-3-甲基咪唑六氟磷酸盐([BMIm][PF6])作为环境选择性压力可以促进抗生素抗性基因的水平转移。本研究以E.coli DH5α(RP4)和同属的E.coli HB101,以及E.coli DH5α(RP4)和跨属的Salmonella enterica之间的纯菌接合转移体系为考察对象,从mRNA基因表达调控水平角度阐明离子液体[BMIm][PF6](0.001~2.5 g·L-1)影响质粒RP4接合转移的机理。结果表明,离子液体[BMIm][PF6]通过抑制基因kor A,kor B和trb A的mRNA表达水平来提高接合和跨膜转运基因trb Bp和trf Ap的表达;增强水平转移基因tra F的mRNA表达水平,并通过增强负调控基因kil A和kil B的mRNA表达水平抑制质粒的垂直传递过程;通过抑制基因trb K的mRNA表达水平降低接合转移体系的排斥效果,从而促进质粒RP4的接合转移。该结果有助于为抗生素抗性基因在环境中水平转移扩散的机理研究提供理论依据。
关键词(KeyWords): 离子液体;抗生素抗性基因;质粒RP4;接合转移;mRNA表达
基金项目(Foundation): 国家自然科学基金(41473085);; 环境污染过程与基准教育部重点实验室开放基金(KL-PPEC-2016-1);; 河北省自然科学基金(No.D2017402109);; 河北省教育厅青年基金(No.QN2017035);; 邯郸市科技计划项目(1621212047)
作者(Author): 汪庆;罗义;崔玉晓;齐丽英;周烁磊;杨光;
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DOI:
参考文献(References):
- [1]Zhu Y G,Zhao Y,Li B,et al.Continental-scale pollution of estuaries with antibiotic resistance genes[J].Nature Microbiology,2017,2:1-7
- [2]Zhu B,Chen Q,Chen S,et al.Does organically produced lettuce harbor higher abundance of antibiotic resistance genes than conventionally produced?[J].Environment International,2017,98:152-159
- [3]苏建强,黄福义,朱永官.环境抗生素抗性基因研究进展[J].生物多样性,2013,21(4):481-487Su J Q,Huang F Y,Zhu Y G.Antibiotic resistance genes in the environment[J].Biodiversity Science,2013,21(4):481-487(in Chinese)
- [4]Pallecchi L,Bartoloni A,Gotuzzo E,et al.Antibiotic Resistance in the Absence of Antimicrobial Use[M]//Microbial Drug Resistance.2013:120-132
- [5]Thomas C M,Nielsen K M.Mechanisms of,and barriers to,horizontal gene transfer between bacteria[J].Nature Reviews Microbiology,2005,3(9):711-721
- [6]Alonso A,Sánchez P,Martinez J L.Environmental selection of antibiotic resistance genes[J].Environmental Microbiology Reports,2001,3(1):1-9
- [7]Wishart J F.Energy applications of ionic liquids[J].Energy Environmental Science,2009,2(9):956-961
- [8]María D D,Maugeri P Z.Ionic liquids in biotransformations:From proof-of-concept to emerging deep-eutecticsolvents[J].Current Opinion in Chemical Biology,2011,15(2):220-225
- [9]Welton T.Room-temperature ionic liquids.Solvents for synthesis and catalysis[J].Chemical Reviews,1999,99(8):2071-2084
- [10]Lu J,Yan F,Texter J.Advanced applications of ionic liquids in polymer science[J].Progress in Polymer Sci-ence,2009,34(5):431-448
- [11]Li Z,Jia Z,Luan Y,et al.Ionic liquids for synthesis of inorganic nanomaterials[J].Current Opinion in Solid State and Materials Science,2008,12(1):1-8
- [12]Pham T P T,Cho C W,Yun Y S.Environmental fate and toxicity of ionic liquids:A review[J].Water Research,2010,44(2):352-372
- [13]Zhao D,Liao Y,Zhang Z.Toxicity of ionic liquids[J].Clean-Soil,Air,Water,2007,35(1):42-48
- [14]Khudyakov J I,Dhaeseleer P,Borglin S E,et al.Global transcriptome response to ionic liquid by a tropical rain forest soil bacterium,Enterobacter lignolyticus[J].Proceedings of the National Academy of Sciences,2012,109(32):2173-2182
- [15]Xue Z,Sendamangalam V R,Gruden C L,et al.Multiple roles of extracellular polymeric substances on resistance of biofilm and detached clusters[J].Environmental Science and Technology,2012,46(24):13212-13219
- [16]Yi L,Wang Q,Lu Q,et al.An ionic liquid facilitates the proliferation of antibiotic resistance genes mediated by class I integrons[J].Environmental Science Technology Letters,2014,1(5):266-270
- [17]Haase J,Lurz R,Grahn A M,et al.Bacterial conjugation mediated by plasmid RP4:RSF1010 mobilization,donor-specific phage propagation,and pilus production require the same Tra2 core components of a proposed DNA transport complex[J].Journal of Bacteriology,1995,177(16):4779-4791
- [18]Jagura-Burdzy G,Thomas C M.kfr A gene of broad host range plasmid RK2 encodes a novel DNA-binding protein[J].Journal of Molecular Biology,1992,225(3):651-660
- [19]Antonenka U,Nlting C,Heesemann J,et al.Horizontal transfer of Yersinia high‐pathogenicity island by the conjugative RP4 att B target‐presenting shuttle plasmid[J].Molecular Microbiology,2005,57(3):727-734
- [20]Llosa M,Gomis-R F X,Coll M,et al.Bacterial conjugation:A two-step mechanism for DNA transport[J].Molecular Microbiology,2002,45(1):1-8
- [21]Jagura-Burdzy G,Khanim F,Smith C A,et al.Crosstalk between plasmid vegetative replication and conjugative transfer:Repression of the trf A operon by trb A of broad host range plasmid RK2[J].Nucleic Acids Research,1992,20(15):3939-3944
- [22]Sakai H,Komano T.DNA replication of Inc Q broadhost-range plasmids in gram-negative bacteria[J].Bioscience Biotechnology and Biochemistry,1996,60(3):377-382
- [23]Lyras D,Chan A W,Mcfarlane J,et al.The surface exclusion system of RP1:Investigation of the roles of trb J and trb K in the surface exclusion,transfer,and slowgrowth phenotypes[J].Plasmid,1994,32(3):254-261
- [24]Haase J,Kalkum M,Lanka E.Trb K.A small cytoplasmic membrane lipoprotein,functions in entry exclusion of the Inc P-alpha plasmid RP4[J].Journal of Bacteriology,1996,178(23):6720-6729
- [25]Chen H,Zhang M M.Occurrence and removal of antibiotic resistance genes in municipal wastewater and rural domestic sewage treatment systems in eastern China[J].Environment International,2013,55:9-14
- [26]Luo Y,Mao D,Rysz M,et al.Trends in antibiotic resistance genes occurrence in the Haihe River,China[J].Environmental Science and Technology,2010,44(19):7220-7225
- [27]Qiu Z G,Yu Y M,Chen Z,et al.Nanoalumina promotes the horizontal transfer of multiresistance genes mediated by plasmids across genera[J].Proceedings of the National Academy of Sciences,2012,109(13):4944-4949
- [28]Suzuki S,Kimura M,Agusa T,et al.Vanadium accelerates horizontal transfer of tet(M)gene from marine Photobacterium to Escherichia coli[J].FEMS Microbiology Letters,2012,336(1):52-56
- [29]Ganske F,Bornscheuer U T.Growth of Escherichia coli,Pichia pastoris and Bacillus cereus in the presence of the ionic liquids[BMIm][BF4]and[BMIm][PF6]and organic solvents[J].Biotechnology Letters,2006,28(7):465-469
- [30]Song X L,Ye S Y,Xie R,et al.Effects of BMIm[PF6]treatments with different concentrations on microbial activity of Saccharomyces cerevisiae[J].Korean Journal of Chemical Engineering,2011,28(9):1902-1907
- [31]Van Hoek A H,Mevius D,Guerra B,et al.Acquired antibiotic resistance genes:An overview[J].Frontiers in Microbiology,2011,3(2):1-17
- [32]Li B,Zhang T.Biodegradation and adsorption of antibiotics in the activated sludge process[J].Environmental Science and Technology,2010,44(9):3468-3473