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Polygene element realizing equivalent expression of two genes based on 2A lytic peptide, expression vector and application

A technology of genetic elements and expression vectors, applied in the direction of vectors, nucleic acid vectors, and the use of vectors to introduce foreign genetic material, etc.

Active Publication Date: 2018-09-04
SOUTHWEST UNIVERSITY
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Problems solved by technology

[0005] However, it has not been reported that the Kozak sequence is used to optimize the 2A multi-gene expression system in the silk gland of silkworm, so that it can recombine and express genes located at both ends of 2A in equal amounts.

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  • Polygene element realizing equivalent expression of two genes based on 2A lytic peptide, expression vector and application
  • Polygene element realizing equivalent expression of two genes based on 2A lytic peptide, expression vector and application
  • Polygene element realizing equivalent expression of two genes based on 2A lytic peptide, expression vector and application

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Embodiment Construction

[0019] The preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings.

[0020] The materials used in the implementation of the present invention are as follows:

[0021] Cell lines: Spodoptera litura Sf9 ovary cell line, silkworm embryo BmE cells. Both Sf9 cells and BmE cells were cultured using medium (Gibco) containing 10% fetal bovine serum (Gibco);

[0022] Target genes: red fluorescent protein gene (DsRed), green fluorescent protein gene (EGFP), and fusion genes DsRed-P2A-CkEGFP, CkEGFP-P2A-CkDsRed and EGFP-P2A-DsRed, all synthesized by GenScript;

[0023] Silkworm strain: non-diapause variety D9L.

[0024] 1. Construction of transient expression vector and transgene expression vector

[0025] According to the design rules of the Kozak sequence: the bases A, U, and G in AUG are marked as 1, 2, and 3 respectively, (1) the preferred base at the 4th position is G; (2) the 5' end of AUG is about There is n...

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Abstract

The invention relates to a polygene element realizing equivalent expression of two genes based on 2A lytic peptide, an expression vector and application. The gene element is formed by Ck and the 2A lytic peptide, wherein a nucleotide sequence of the 2A lytic peptide is as shown in SEQ ID NO.38, and a nucleotide sequence of the Ck is as shown in SEQ ID NO.39. The vector containing the element can be used for equivalently performing recombination expression on the genes located on two ends of 2A so as to have significance on realizing silkworm poly-gene expression.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a multigene element based on 2A cleavage peptide for realizing the equal expression of two genes, and also relates to an expression vector containing the element and its application. Background technique [0002] Efficient recombinant expression of multiple genes in silkworms can promote basic and applied research in silkworms, such as studying the relationship between multiple genes, recombinantly expressing genes of multi-subunit protein complexes, and simultaneously expressing fluorescent proteins to visualize targets The movement of genes in the body and the breeding of silkworm varieties resistant to silkworm cytoplasmic polyhedrosis virus and nuclear polyhedrosis virus, etc. Thanks to the establishment of the silkworm transgenic operating system in 2000, previously, simultaneous expression of multiple genes in the silkworm was mainly achieved through genetic hybridization. First...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/85
CPCC12N15/113C12N15/85C12N2800/105C12N2840/44
Inventor 王峰夏庆友王元成王日远陈文静赵萍
Owner SOUTHWEST UNIVERSITY
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