Bicistronic mRNA coexpression gene transporter and preparation method thereof

Abstract

The invention relates to a bicistronic mRNA coexpression gene transporter and a preparation method thereof. The gene order of the gene transporter is SEQ ID No. 15; the gene transporter from 5' end to 3' end comprises bovine Nuclear matrix binding region MAR, artificially constructed combined promoter CAG, Profilin gene FSTN, internal ribosome entry site (IRES), green fluorescent protein AcGFP gene and Rabbit globin poly A signal region; the preparation method comprises the following steps: constructing a carrier pCAG-IRES2-AcGFP1; obtaining FSTN gene and inserting the pCAG-IRES2-AcGFP1 carrier; obtaining the sequence of MAR and inserting the pCAG-IRES2-AcGFP1 carrier; constructing a plasmid carriers so as to obtain the bicistronic mRNA coexpression gene transporter. The gene transporter is not only pure and safe gene transporter but also realizes dual-gene coexpression in any combination, achieves the purpose of multi-gene coexpression through repeated utilization of IRES, and provides new thinking and route for improving the shape of dual or multiple gene control such as economic character.

Description

technical field [0001] The invention belongs to the genetic engineering technology in the field of biotechnology, and in particular relates to a preparation method of a bicistronic co-expression gene transfer body. Background technique [0002] High-efficiency expression of foreign genes in transgenic animals must rely on good expression vectors. There are many factors that affect the high-efficiency expression of foreign genes, such as promoters, methylation, gene structure, insertion sites, regulatory sequences (enhancers, insulators, nuclear matrix binding regions), etc. [0003] The CAG promoter is an artificially constructed combined promoter consisting of the early enhancer element of the cytomegalovirus (CMV) and the chicken beta-actin promoter. The CAG promoter is Nonspecific constitutive promoters for driving high-level expression of genes in mammalian vectors. CMV is a commonly used promoter. In the application of our transgenic sheep, it was found that methylati...

AI Technical Summary

Problems solved by technology

[0009] In recent years, the gene expression cassette (including only the promoter, coding region and terminator) with the backbone sequence of the plasmid vector removed has been used as a gene transfer body, also known as clean DNA transformation (clean DNA transformation), mainly through the transformation of plants by gene guns, and has been used in rice. It has been successfully applied in the transformation of cotton, wheat, and grapes, and it has also been successfully applied in muskmelon by using the pollen tube introduction method. It is rarely used in the construction and application of transgenic animals, especially mammals.

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