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A kind of pkd46-arr-3 plasmid and its preparation method

A pkd46-arr-3, plasmid technology, applied in the direction of recombinant DNA technology, introduction of foreign genetic material using vectors, etc.

Active Publication Date: 2016-07-06
THE SECOND HOSPITAL AFFILIATED TO SUZHOU UNIV
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  • Abstract
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Problems solved by technology

The screening tag of pKD46 is the ampicillin resistance gene, but the natural ampicillin resistance of clinical strains is very common, especially the ampicillin natural resistance of some Gram-negative Enterobacteriaceae is particularly common, which limits the use of the pKD46 plasmid in these ampicillin-resistant Application in resistant strains

Method used

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  • A kind of pkd46-arr-3 plasmid and its preparation method
  • A kind of pkd46-arr-3 plasmid and its preparation method
  • A kind of pkd46-arr-3 plasmid and its preparation method

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Embodiment Construction

[0027] The above solution will be further described below in conjunction with specific embodiments. It should be understood that these examples are used to illustrate the present invention and not to limit the scope of the present invention. The implementation conditions used in the examples can be further adjusted according to the conditions required by specific applications, and the unspecified implementation conditions are usually the conditions in routine experiments.

[0028] Materials and Instruments

[0029] BssSI and PvuI endonuclease, Taq enzyme, T4 ligase were all purchased from Takara Company;

[0030] Electrophoresis grade agarose, yeast extract, tryptone, agar powder, etc. were purchased from Sangon Bioengineering (Shanghai) Co., Ltd.

[0031] The pKD46 plasmid was purchased from Wuhan Miaoling Biotechnology Co., Ltd. Klebsiella pneumoniae was purchased from Nanjing Bianjian Biotechnology Co., Ltd.

[0032] PCR instrument - BioRad (USA), gel imaging system - B...

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Abstract

The invention relates to the field of molecular biology, and discloses a plasmid pKD46-arr-3 and a preparation method thereof. The plasmid comprises a pKD46 carrier gene segment and a rifampicin resistance gene arr-3. The plasmid pKD46-arr-3 is used for removing a target gene by introducing ammonia benzyl resistant escherichia coli, klebsiella pneumoniae and other strains clinically separated.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to molecular biology and genetic engineering. Background technique [0002] Plasmid is an extrachromosomal genetic unit capable of self-replicating. It exists widely in many organisms. It is customarily used to refer to DNA molecules outside the chromosome in organisms such as bacteria, yeast, and actinomycetes. It is also commonly used in molecular biology techniques. a tool molecule. [0003] The pKD series vectors mainly include pKD3, pkD4, pKD13, pKD32 and pKD46, etc., which are tool plasmids used to prepare bacterial gene-deficient strains through homologous recombination, among which pKD46 is mainly used for gene knockout of Gram-negative bacilli. The screening tag of pKD46 is the ampicillin resistance gene, but the natural ampicillin resistance of clinical strains is very common, especially the ampicillin natural resistance of some Gram-negative Enterobacteriaceae is particularl...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/70C12N15/74C12N15/66
Inventor 张海方杜鸿王敏朱雪明谢小芳周惠琴郑毅
Owner THE SECOND HOSPITAL AFFILIATED TO SUZHOU UNIV
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