Expression vector for animal cell as well as preparation method and application of expression vector

Abstract

The invention relates to the field and particularly relates to an expression vector for an animal cell. The expression vector sequentially contains a promoter with strong expression inductivity, a multiple cloning site for gene integration, a ribosome entry site sequence, a dihydrofolate reductase gene, a SV40 early stage promoter gene without an enhanser, a neo gene and an anti-drug gene from upstream. The invention further provides a preparation method of the expression carrier, a recombinant carrier and application. According to the expression carrier provided by the invention, expression of the neo gene is weakened by using the promoter without the promoter; a dhfr gene as a co-amplification gene is located at the downstream of the ribosome entry site IRES, so that the translation efficiency of the dhfr gene is reduced; the expression quantity of a target protein is increased by combining G418 pressurized screening and pressure screening of the dihydrofolate reductase gene.

Description

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AI Technical Summary

Benefits of technology

The technical effects of this patented technology include improved production levels by adding specific genes or parts thereto during cell culture processes instead of relying on other methods like increasing methionine synthesis (which can be expensive). This results from utilizing a strong promoter for driving the transcription process rather than just making it harder to produce new proteins due to its low activity level compared to existing techniques such as cloning.

Problems solved by technology

Technologies described involve improving the ability of producing highly effective amounts of desired products from cloned animal tissue sources called germ line fractions through fusion techniques involving specific parts of the parent molecule's own genomes. By combining multiple different components like other gene segments within certain regions of the recipients, even if some genes were deleted during culture process, the transformed product could remain stable overtime without losing any key structural features found naturally occurring ones. Additionally, modifying the expression rate of each component allows for efficient expression of various genes while minimizing changes caused by external influences including pigment accumulation, oxidative stress, and reduced sensitivity due to decreased transferring efficacy. Overall, these technical means improve the quality and quantity of produced materials derived from CHOs cells.

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