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无痕敲除法构建食品安全级枯草芽孢杆菌无芽孢菌株

Construction of a food safe grade Bacillus subtilis nonspore strain with clean deletion
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摘要 利用同源重组法快速构建枯草芽孢杆菌(Bacilus subtilis168)spo0A基因缺陷型无芽孢菌株B.subtilis168Δspo0A。经PCR及核酸电泳验证,孢子形成率鉴定和芽孢染色镜检,确定最终获得不产芽孢的B.subtilis168Δspo0A基因缺失突变菌株。基因敲除完成后,菌株基因组不需引入抗性基因或其它任何筛选标记,实现了对spo0A基因的无痕基因敲除。本研究对枯草杆菌的工业化生产及提高相关产品食品质量安全性具有重要意义。 Construction a nonspore B.subtilis 168 strain( B.subtilis 168Δspo0A) by deleting the spo0 A gene using the method of homologous recombination.The nonspore B.subtilis168Δspo0A strain was successfully got through PCR,nucleic acid electrophoresis,sporulation ratio and microscopy identification.The genome of this engineering strain does not need to insert resistance gene or any other selection marker gene after spo0 A genes' knockout which called clean deletion. This study is of great significance for industrial production of B.subtilis as well as improve safety of food and drug.
出处 《食品工业科技》 CAS CSCD 北大核心 2016年第14期175-180,共6页 Science and Technology of Food Industry
关键词 枯草芽孢杆菌(Bacilus subtilis168) 无痕基因敲除 无芽孢菌株 Bacilus subtilis168 clean deletion nonspore strain
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参考文献12

  • 1Wang ZW, Chen T, Ma XH, el al. EnhaneemenI of riboflavin production with Bacillus subtilis by expression and site-directed nmtagenesis of zuf and gml gene from Corynebacterium glutamicum [J] . Bioresouree Technology, 2011, 102 ( 4 ) : 3934 -3940.
  • 2Phan 'rI'P, Nguyen HD, Schumann W. Constntction of a 5 '-controllable stabilizing element ( CoSE ) t)r over- production of heterologous proleins at high levels in Bwillus subtilis [J] .J Bioteehnology, 2013,168 ( 1 ) : 32- 39.
  • 3Wu L, Li Z, Ye Q.Enhanced d-ribose biosynthesis in batch culture of a transketolase- deficient Bacillus subtilis strain hy citrate[J] .Journal of Industrial Microbiology & Bioteehnology, 2009,36(10) :1289-1296.
  • 4Sehallmey M,Singh A, Ward OP.Developments in the use of Bacillus species for induslrial production [J] .Canadian Journal of Mic.robiology, 2004,50 ( 1 ) : 1 - 17.
  • 5沈卫锋,牛宝龙,翁宏飚,何丽华,孟智启.枯草芽孢杆菌作为外源基因表达系统的研究进展[J].浙江农业学报,2005,17(4):234-238. 被引量:25
  • 6刘燕,秦玉昌,潘宝海.枯草芽孢杆菌(Bacillus subtilis)在芽孢形成过程中的几个关键事件[J].生命科学,2005,17(4):360-363. 被引量:10
  • 7Rowe-Magnus DA, Spiegelman GB.DNA strand separation during activation of a developmental promoter by the Bacillus subtilis response regulator spo0A[J] .Proceedings of the National Aeademy of Sciences of the United States of America, 1998,95 (9) :5305-5310.
  • 8余志强,杨明明,杨朝霞,林峰,张西锋,张炜炜,樊俊华,姜发堂.同源重组法构建枯草杆菌spo0 A基因缺失突变株[J].武汉大学学报(理学版),2004,50(2):229-233. 被引量:7
  • 9Maryvonnc Arnaud, Arnaud Chastanct, Michel Debarbouille.New vector for effMent allelic replacement innatually nontransformable, low- GC - content, gram- positive bacteria [J] .Applied & Environmental Microbiology, 2004, 70 ( 11 ) :6887-6891.
  • 10甄静,郭直岳,谢宝恩,李冠杰,刘莹莹,慕琦.枯草芽孢杆菌XK-1产芽孢条件的优化[J].中国农学通报,2012,28(27):146-151. 被引量:24

二级参考文献73

  • 1陈乃用.枯草芽孢杆菌中质粒的稳定性问题[J].微生物学通报,1993,20(4):226-232. 被引量:8
  • 2葛诚,吴薇.我国微生物肥料的生产、应用及问题[J].中国农学通报,1994,10(3):24-28. 被引量:58
  • 3郭小华,陆文清,邓萍,黄德仕.益生枯草芽孢杆菌MA139增殖培养基的优化[J].中国农业大学学报,2006,11(3):41-46. 被引量:25
  • 4宋卡魏,王星云,张荣意.培养条件对枯草芽孢杆菌B68芽孢产量的影响[J].中国生物防治,2007,23(3):255-259. 被引量:33
  • 5Marchioli C C,Yancey R J Jr,Wardley R C,et al.A vaccine strain of pseudorabies virus、with deletions in the thymidine kinase and glycoprotein X genes[J].Am J Vet Res,1987,48(11):1577-1583.
  • 6Lisa E,Pomeranz A,Shley E,et al.Molecular Biology of Pseudorabies Virus:Impact on Neurovirology and Veterinary Medicine[J].Microbiol Mol Biol Rev.2005,69(3):462-500.
  • 7Jacobs L,Rziha H J,Kimman T G,et al.Deleting valine-125 and cysteine-126 in glycoprotein gI of pseudorabies virus strain NIA-3 decreases plaque size and reduces virulence in mice[J].Arch Virol,1993,13l(3-4):251-264.
  • 8Quint w,Gielkens A,Van Van Oirschot J,et al.Construction and Characterization of Deletion Mutants of Pseu dorabies Vims:a New Generation of‘Live'Vaccines[J].J Gen Virol,1987,68:523-534.
  • 9Song Y F,Jin M L,Zhang S L,et al.Generation and immunogenicity of a recombinant pseudorabies virus expressing cap protein of porcine circovirus type 2[J].Vet Microbiol,2007,119(1):97-104.
  • 10Jacobs L,Rziha H J,Kimman T G,et al.Glycoprotein I of pseudorabies virus(Aujeszky's disease virus)determines virulence and facilitates penetration of the virus into the central nervous system of pigs[J].Acta Vet Hung,1994,42(2-3):289-300.

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