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Plasmid pKD46-arr-3 and preparation method thereof

A technology of pkd46-arr-3 and plasmids, which is applied in the direction of recombinant DNA technology, the use of vectors to introduce foreign genetic material, etc.

Active Publication Date: 2015-11-11
THE SECOND HOSPITAL AFFILIATED TO SUZHOU UNIV
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  • Abstract
  • Description
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AI Technical Summary

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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  • Plasmid pKD46-arr-3 and preparation method thereof
  • Plasmid pKD46-arr-3 and preparation method thereof
  • Plasmid pKD46-arr-3 and preparation method thereof

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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 embodiments 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 the specific application, and the implementation conditions not specified are usually the conditions in the routine experiment.

[0028] Materials and instruments

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

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

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

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

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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 autonomous replication. It is widely present in many organisms. It is customarily used to specifically refer to DNA molecules other than chromosomes in organisms such as bacteria, yeasts, and actinomycetes. It is also commonly used in molecular biology techniques. A kind of tool molecule. [0003] The pKD series of 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 them, pKD46 is mainly used for gene knockout of gram-negative bacilli. The screening label of pKD46 is the ampicillin resistance gene, but the natural resistance of ampicillin in clinical strains is very common, especially the natural resistance of ampicillin in some Gram-negat...

Claims

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Application Information

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Patent Type & Authority Applications(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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