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A new cloning vector pat3733

A technology of pat3733 and cloning vector, applied in the field of cloning vector plasmid, can solve the problems of low DNA copy number, low yield, affecting the efficiency and effect of experimental research, and achieve the effect of small molecular weight

Inactive Publication Date: 2016-01-06
YUNNAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In pBR322, due to Rop Due to the existence of genes, the replication of plasmid DNA is seriously affected, the copy number of DNA is low, and the yield is low, which affects the efficiency and effect of experimental research

Method used

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  • A new cloning vector pat3733

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Embodiment Construction

[0013] The biological material pBR322 involved in the present invention is a published or commercialized material, specifically purchased from Takara Company.

[0014] In the embodiment shown in the accompanying drawings, the 86th-1276bp is a tetracycline resistance gene with a size of 1190bp, wherein Eco RV (187bp), Bam HI (375bp), Sal I (651bp) and other restriction endonuclease cutting sites. 1891-2505bp is rep Gene, the size is 614bp, which has Apa LI (2159bp), Bfa I (2340bp) and other restriction endonuclease cutting sites. No. 2665-3525 is the ampicillin resistance gene, the size is 860bp, and there are Bgl I (2858), Pst I (2963bp), Pvu I (3108) and other restriction endonuclease cutting sites. The size of the whole plasmid is 3733bp.

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Abstract

The invention relates to an insertional inactivation type cloning vector pAT 3733 with ampicillin and tetracycline resistant genes. The vector is obtained by the following steps: removing the Rop gene on pBR322 by double digestion method, filling in the end and looping internally. pAT3733 not only preserves the insertional inactivation screening mark of pBR322, but also has smaller molecular weight and higher stability, and the plasmid copy number increases more than three times, and the product is convenient for gene clone operation with higher operating efficiency. The product can replace pBR322 plasmid which is widely used at present, and is used as a general high efficiency cloning vector.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a cloning carrier plasmid. Background technique [0002] Plasmid pBR322 is an ideal cloning vector that was first successfully constructed by artificial methods. It is still the most commonly used and most representative plasmid in genetic engineering. It is widely used in genetic engineering and is known as the "universal vector". It was constructed by Bolivar et al. in 1977 and is a typical artificial plasmid vector. It has a small relative molecular mass, so it loads a large amount of foreign DNA; it has a high copy number, which provides great convenience for the preparation of recombinant DNA; it also has two antibiotic resistance genes , namely ampicillin resistance gene and tetracycline resistance gene. This provides convenience for the detection of recombinant plasmids. However, the ROP (RepressorOfPrimer) factor in the pBR322 plasmid controls the copy number of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/63
Inventor 韦建福
Owner YUNNAN AGRICULTURAL UNIVERSITY
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