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  • 标题:Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti
  • 本地全文:下载
  • 作者:G Ditta ; S Stanfield ; D Corbin
  • 期刊名称:Proceedings of the National Academy of Sciences
  • 印刷版ISSN:0027-8424
  • 电子版ISSN:1091-6490
  • 出版年度:1980
  • 卷号:77
  • 期号:12
  • 页码:7347-7351
  • DOI:10.1073/pnas.77.12.7347
  • 语种:English
  • 出版社:The National Academy of Sciences of the United States of America
  • 摘要:A broad host range cloning vehicle that can be mobilized at high frequency into Gram-negative bacteria has been constructed from the naturally occurring antibiotic resistance plasmid RK2. The vehicle is 20 kilobase pairs in size, encodes tetracycline resistance, and contains two single restriction enzyme sites suitable for cloning. Mobilization is effected by a helper plasmid consisting of the RK2 transfer genes linked to a ColE1 replicon. By use of this plasmid vehicle, a gene bank of the DNA from a wild-type strain of Rhizobium meliloti has been constructed and established in Escherichia coli. One of the hybrid plasmids in the bank contains a DNA insert of approximately 26 kilobase pairs which has homology to the nitrogenase structural gene region of Klebsiella pneumoniae.
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