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流产布鲁氏菌的转座子Tn5诱变

Transposon Tn5 mutagenesis of Brucella abortus.

作者信息

Smith L D, Heffron F

机构信息

Department of Molecular Biology, Research Institute of Scripps Clinic, La Jolla, California 92037.

出版信息

Infect Immun. 1987 Nov;55(11):2774-6. doi: 10.1128/iai.55.11.2774-2776.1987.

DOI:10.1128/iai.55.11.2774-2776.1987
PMID:2822580
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC259975/
Abstract

We demonstrate that transposon-mediated mutagenesis can be used to construct mutations in the pathogen Brucella abortus. We have used both a plasmid and a bacteriophage to introduce either Tn5 or Tn5 lac into the Brucella chromosome. B. abortus is naturally sensitive to kanamycin. We have selected 22 independent kanamycin-resistant colonies in strain US-19 and 19 colonies in strain 2308. We have demonstrated by Southern hybridization that Tn5 was inserted into the chromosome.

摘要

我们证明转座子介导的诱变可用于在流产布鲁氏菌病原体中构建突变体。我们使用了质粒和噬菌体将Tn5或Tn5 lac导入布鲁氏菌染色体。流产布鲁氏菌对卡那霉素天然敏感。我们在US-19菌株中筛选出22个独立的卡那霉素抗性菌落，在2308菌株中筛选出19个菌落。我们通过Southern杂交证明Tn5已插入染色体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d78f/259975/035e6793d740/iai00095-0246-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d78f/259975/035e6793d740/iai00095-0246-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d78f/259975/035e6793d740/iai00095-0246-a.jpg

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本文引用的文献

1
Differentiation of smooth and nonsmooth colonies of Brucellae.布鲁氏菌光滑型和粗糙型菌落的鉴别
J Bacteriol. 1951 Feb;61(2):239-40. doi: 10.1128/jb.61.2.239-240.1951.
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Deletion of the BCSP31 gene of Brucella abortus by replacement.通过置换缺失流产布鲁氏菌的BCSP31基因。
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Construction of Tn5 lac, a transposon that fuses lacZ expression to exogenous promoters, and its introduction into Myxococcus xanthus.构建将lacZ表达与外源启动子融合的转座子Tn5 lac，并将其导入黄色粘球菌。
Proc Natl Acad Sci U S A. 1984 Sep;81(18):5816-20. doi: 10.1073/pnas.81.18.5816.
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Bacteriophage receptors.噬菌体受体
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Analysis of endonuclease R-EcoRI fragments of DNA from lambdoid bacteriophages and other viruses by agarose-gel electrophoresis.通过琼脂糖凝胶电泳分析来自λ样噬菌体和其他病毒的DNA的核酸内切酶R-EcoRI片段。
J Virol. 1974 Nov;14(5):1235-44. doi: 10.1128/JVI.14.5.1235-1244.1974.
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Shuttle mutagenesis: a method of transposon mutagenesis for Saccharomyces cerevisiae.穿梭诱变：一种用于酿酒酵母的转座子诱变方法。
Proc Natl Acad Sci U S A. 1986 Feb;83(3):735-9. doi: 10.1073/pnas.83.3.735.
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Restriction endonuclease analysis of Brucella ovis and other Brucella species.绵羊布鲁氏菌及其他布鲁氏菌属菌种的限制性内切酶分析。
Vet Microbiol. 1985 Aug;10(5):425-9. doi: 10.1016/0378-1135(85)90024-0.
9
A restriction enzyme cleavage map of Tn5 and location of a region encoding neomycin resistance.Tn5的限制性内切酶切割图谱及编码新霉素抗性区域的位置。
Mol Gen Genet. 1979;177(1):65-72. doi: 10.1007/BF00267254.