A bicistronic expression vector suitable for hek293 cells and its preparation method, expression system, and application

1. HEK293, expression system technology, applied in the direction of using vectors to introduce foreign genetic material, recombinant DNA technology, etc., can solve the problems of inability to screen stably transformed cell lines, not including screening markers, etc.
CN107058387BActive Publication Date: 2019-09-06河南普诺易生物制品研究院有限公司 +1
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
河南普诺易生物制品研究院有限公司
Publication Date
2019-09-06

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Abstract

The invention relates to a two-cistron expression vector suitable for an HEK293 cell, and a preparation method, an expression system and application thereof, which belong to the technical field of genetic engineering. The two-cistron expression vector suitable for an HEK293 cell comprises the following elements: a first nuclear matrix combined region sequence-promoter-target gene-internal ribosome entry site sequence-screened marker genes-poly A-second nuclear matrix combined region sequence. According to the vector, by utilizing the optimized MAR sequence, the transgenic expression of the HEK293 cell can be significantly improved, the transgenic silencing is overcome, and the efficient long-term expression of the transgene in a host cell can be realized. Meanwhile, the simultaneous expression of the target gene and the screened marker genes can be realized, the false positive cell clone caused by the existence of different expression cassettes can be reduced, the positive cell clone screening rate is increased, and the problem of imbalanced expression of the target gene and the screened marker gene of the traditional vector can be solved.
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Description

technical field

[0001] A bicistronic expression vector suitable for HEK293 cells and its preparation method, expression system and application belong to the technical field of genetic engineering. Background technique

[0002] The development of genetic engineering technology in the 1970s transformed the use of animal organs, tissues or human blood to extract protein drugs into the production of genetic engineering. Pharmaceutical proteins, including monoclonal antibodies, peptides, and proteins, have become an important part of the pharmaceutical industry and will become an important area of ​​future pharmaceutical development.

[0003] The expression systems of recombinant drug proteins mainly include E.coli, yeast and non-humanized mammalian cell lines. E.coli is suitable for expressing proteins with small molecular weight and relatively simple structure, and the yeast expression system is suitable for expressing proteins with large molecular weight, complex structure an...

Claims

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