期刊文献+
任意字段
题名或关键词
题名
关键词
文摘
作者
第一作者
机构
刊名
分类号
参考文献
作者简介
基金资助
栏目信息

鳗弧菌Vibro anguillarum MVM425sh高效电转化条件的优化 被引量：1

Optimization of Technical Condition for Transformation of Vibro anguillarum MVM425sh Strain by Electroporation
在线阅读 下载PDF
导出
摘要 目的建立高效的鳗弧菌电转化系统。方法分别从感受态细胞制备的洗涤缓冲液与浓度、复苏培养基、二级细胞收获时间、电转化的电压与脉冲时间等方面,对鳗弧菌的电转化条件进行优化。结果获得的鳗弧菌最佳转化条件为二级培养3h左右收获细胞,用272mmol/L蔗糖洗涤缓冲液制备电转化细胞,在2.0kV3ms的脉冲条件下,pUC18的转化率达106个转化子/μgDNA。结论采用优化的电转化方法,可获得较高的转化率,为鳗弧菌中的常规遗传操作奠定了良好的基础。 Objective To optimize the technical condition for transformation of Vibro anguillarum MVM425sh strain by electroperation. Methods Explore the influence of technical conditions such as washing solution of competent cells, medium for cell resuscitation, recovery time of secondary cells, voltage for transformation and pulse time on transformation efficay of Vibro anguillarum MVM425 sh strain. Results The condition for transformation was optimized as follows：recover cells 3 h after secondary culture, prepare electrotransformed cells using 272 mmol/L sucrose as washing solution and transform competent cells at 2. 0 kV for 3 ms. The transformation efficacy under this condition reached 106 transformants/μg DNA. Conclusion High transformation efficacy was obtained under the optimized condition. The study laid a foundation of routine genetic manipulation of Vibro anguillarum.
作者 吴海珍 张惠展 梁娜 顾振霆 叶江 张元兴
机构地区 华东理工大学生物反应器工程国家重点实验室
出处 《中国生物制品学杂志》 CAS CSCD 2007年第8期600-602,608,共4页 Chinese Journal of Biologicals
关键词 鳗弧菌 电转化 质粒 转化率 Vibro anguillarum Electroporation Plasmid Transformation efficacy
分类号 Q7 [生物学—分子生物学]
  • 相关文献

参考文献8

  • 1Lorenzo MD,Stork M,Tolmasky ME,et al.Complete sequence of virulence plasmid pJM1 from the marine fish pathogen Vibrio anguillarum strain 775.J Bacteriol,2003,185(19):5822-5830.
  • 2Wu H,Ma Y,Zhang Y,et al.Complete sequence of virulence plasmid pEIB1 from the marine fish pathogen Vibrio anguillarum strain MVM425 and location of its replication region.J Appl Microbiol,2004,97(5):1021-1028.
  • 3Strom MS,Paranjpye RN.Epidemiology and pathogenesis of Vibrio vulnificus.Microbes Infect,2000,2(2):177-188.
  • 4Cutrin JM,Toranzo AE,Barja JL.Genetic transformation of Vibrio anguillarum and Pasteurella piscicida by electroporation.FEMS Microbiol Lett,1995,128(11):75-80.
  • 5Liu Q,Ma Y,Wu H,et al.Cloning,identification and expression of an entE homologue angE from Vibrio anguillarum serotype O1.Arch Microbio,2004,181(4):287-293.
  • 6Marcus H,Ketley J,Kaper JB,et al.Effects of Dnase production,plasmid size,and restriction barriers on transformation of Vibrio cholerae by electroporation and osmotic shock.FEMS Microbiol Lett,1990,56(1-2):149-154.
  • 7Serror P,Saski T,Ehrlich SD,et al.Electrotransformation of Lactobacillus delbrueckii subsp.bulgaricus and L.delbrueckii subsp.lactis with various plasmids.Appl Environ Microbiol,2002,68(1):46-52.
  • 8Kim YH,Han KS,Oh S,et at.Optimization of technical conditions for the transformation of Lactobacillus acidophilus strains by electroporation.J Appl Microbiolo,2005,99(1):167-174.

同被引文献21

  • 1Thune R L,Hawke J P,Sibeling R J. Vibriosis in the red swamp crawfish [ J ]. Journal of Aquatic Animal Health,1991,3(3) :97 - 101.
  • 2Chitov T, Kirikaew P, Yungyune P, et al. An incidence of large foodborne outbreak associated with Vibrio mimicus [J]. European Journal of Clinical Microbiology. 2009,28 ( 4 ) :421 - 424.
  • 3Jonas R, Charlotte K, Evgenia P, et al. A simple, versatile method for GFP-based single molecule localization microscopy [ J ]. Biophysical Journal, 2012,102(3 Suppl 1) :419a.
  • 4Baker S M,Buckheit R W, Falk M M. Green-to-red photoconvertible fluorescent proteins: Tracking cell and protein dynamics on standard wide-field mercury arc-based microscopes[J]. BMC Cell Biology,2010, 11(1):1-10.
  • 5Ramsden R, Arms L,Davis T N, et al. An intein with genetically selectable markers provides a new approach to internally /abel proteins with GFP [J]. BMC Biotechnology, 2011,11 ( 1 ) : 71 - 81.
  • 6Shimomura O. Structure of the chromophore of Aequorea green fluorescent protein [ J ]. FEBS Letters, 1979,104 ( 2 ) :220 - 222.
  • 7Yu TT, Skinner D Z, Liang G H, etal. Agrobacterium-mediated transformation of creeping bentgrass using GFP as a reporter gene [J]. Hereditas ,2004,133 ( 3 ) :229 - 233.
  • 8Chu W H, Lu C P. In vivo fish models for visualizing Aeromonas hydrophila invasion pathway using GFP as a biomarker[J]. Aquaculture,2008,277 (3 -4 ) : 152 - 155.
  • 9Aki N, Nobuhiro M, Hitomi H, et al. Localization analysis of fluorescent-labeled Aeromonas veronii in the intestinal tract of carp using an in vivo imaging system [J]. Fish Pathology ,2012,47 ( 1 ) :7 - 11.
  • 10O' Toole R, Von H J,Rosqvist R, et al. Visualisation of zebrafish infection by GFP-labelled Vibrio anguillarum [J].Microbial Pathogenesis, 2004, 37 (1):41 -46.

引证文献1

二级引证文献6

中国生物制品学杂志
2007年 第8期

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部