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Efficient mammalian cell gene recombination protein expression vector

A protein expression and mammalian technology, applied in the field of high-efficiency expression of gene recombinant protein, can solve the problem of low expression

Inactive Publication Date: 2012-11-21
朱燕华
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Problems solved by technology

[0003] In order to overcome the deficiency of the low expression level of the existing mammalian cell gene recombinant protein expression vector, the present invention provides a mammalian cell gene recombinant protein expression vector, which can not only Cell gene recombination protein expression vectors express target proteins in mammalian cells, and the expression efficiency is much higher than these vectors, and its protein yield is 3-50 times that of these vectors

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  • Efficient mammalian cell gene recombination protein expression vector
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Embodiment Construction

[0008] exist figure 1 Among them, the newly inserted DNA fragment (1) with a length of 363 base pairs contains a large number of binding sites for transcription factors, and transcription factors that enhance transcription, such as SP, AP, and CREB / CBP, can bind to the corresponding binding sites Combined with the pCMV promoter (2) to exert a transcriptional enhancement effect and increase protein expression. The cloning site (3) is mainly used to insert the cDNA fragment of the target protein. SV40 pA(4) is a polyA tailing signal required for mRNA processing of eukaryotic genes. SV40 early promoter (5), neomycin / kanamycin resistance gene (6) and polyadenine signal of TK gene (7) constitute the Neo resistance cassette, which can be used to screen stably transfected eukaryotic cell lines with G418. In addition, the pUC origin in the vector can ensure the replication of the vector in E. coli, and the bacterial promoter located upstream of this expression cassette can drive the...

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Abstract

The invention relates to an efficient mammalian cell gene recombination protein expression vector, wherein expression of target protein in mammalian cells by the existing commonly-used mammalian cell gene recombination protein expression vectors can be performed by the efficient mammalian cell gene recombination protein expression vector of the present invention, and the efficient mammalian cell gene recombination protein expression vector of the present invention provides higher expression efficiency compared with the existing vectors. According to the efficient mammalian cell gene recombination protein expression vector of the present invention, a section of DNA sequence with a length of 363 base pairs is inserted into a position at the upstream of a promoter of a vector, wherein the DNA sequence contains a large number of transcription factor binding sites, transcription factors such as SP, AP, CREB / CBP, and the like can bind with the corresponding binding sites so as to provide a transcription enhancement effect and increase a protein expression level, wherein the transcription factors such as SP, AP, CREB / CBP, and the like provide enhancement effects for transcription. The DNA fragment-containing expression vector is introduced into mammalian cells, and the DNA fragment can attract a large number of transcription factors such as SP, AP, CREB / CBP, and the like, such that an expression level of the target protein can be increased by 3-50 times so as to achieve a purpose of protein yield improvement.

Description

technical field [0001] The invention relates to a gene carrier for expressing gene recombination protein in mammalian cells, especially capable of expressing gene recombination protein in mammalian cells with high efficiency. Background technique [0002] Since the birth of genetic engineering technology in the 1970s, gene expression technology has penetrated into various fields of life science research. Using expression vectors to express target genes in mammalian cells is an important means to study gene functions and their interactions, and it is also the main means used by the biopharmaceutical industry to produce gene recombinant protein drugs and antibody drugs. Mammalian cell gene recombination protein expression vector is a DNA segment including components such as prokaryotic sequence, promoter, enhancer, selection marker gene, terminator and polynucleotide signal. The main method of this technology is to introduce the vector that has been cloned into the DNA fragme...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85
Inventor 朱燕华
Owner 朱燕华
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