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Expression cassette, recombinant plasmid and application of skin-specific expression of sheep β-catenin gene

A gene and skin technology, which is applied to the expression cassette, recombinant plasmid and their application fields that specifically express the sheep β-catenin gene in the skin, and can solve the problems that the mode and mechanism of β-catenin are not very clear.

Active Publication Date: 2016-02-24
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the mode and mechanism of action of β-catenin in the process of hair follicle formation and cycle change are still unclear

Method used

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  • Expression cassette, recombinant plasmid and application of skin-specific expression of sheep β-catenin gene
  • Expression cassette, recombinant plasmid and application of skin-specific expression of sheep β-catenin gene
  • Expression cassette, recombinant plasmid and application of skin-specific expression of sheep β-catenin gene

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Experimental program
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Embodiment 1

[0026] Embodiment 1, construction and application of plasmid

[0027] 1. Construction of a recombinant plasmid for skin-specific expression of sheep β-catenin gene

[0028] 1. Genomic DNA extraction from human blood tissue.

[0029] 2. Using the genomic DNA in step 1 as a template, perform PCR amplification with a primer pair composed of K14F and K14R to obtain a PCR amplification product.

[0030] K14F: 5′-C ATTAAT ATCCCTGCAGAAGAAGGAGAC-3';

[0031] K14R: 5′-ATA AGCGCT GGCTGAGTGAAGAGAAGG-3'.

[0032] PCR reaction program: pre-denaturation at 95°C for 5 minutes; denaturation at 95°C for 30 s, annealing at 57°C for 30 s, extension at 72°C for 2 min, 30 cycles; extension at 72°C for 10 min.

[0033] The 1% agarose gel electrophoresis of the PCR amplification product is shown in figure 2 . Specific bands around 2000bp can be observed.

[0034] 3. Double-digest the PCR amplification product of step 2 with restriction endonucleases Asel and AfeI, and recover the digested...

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Abstract

The invention discloses an expression cassette for specifically expressing a sheep beta-catenin gene in skin, a recombinant plasmid and application of the expression cassette and the recombinant plasmid. The invention provides a deoxyribonucleic acid (DNA) molecule. The DNA molecule comprises a human skin specific promoter K14 and a beta-catenin gene sequentially from upstream to downstream in sequence. Beta-catenin gene protein is as shown in the sequence 2 of a beta-catenin gene coding sequence table. The human skin specific promoter K14 is specifically shown as the 12 to the 2011 nucleotide of the 5' terminal of the sequence 1 of the sequence table. The invention provides the corresponding expression cassette and the recombinant plasmid, which can specially drive the beta-catenin gene to express in animal skin and promote growth of skin hair follicle so as to improve the wool yield and the wool quality of the animal on the premise of not damaging the normal growth of the individual.

Description

technical field [0001] The invention relates to an expression cassette for specifically expressing sheep β-catenin gene in skin, a recombinant plasmid and applications thereof. Background technique [0002] The hair follicle is a complex skin accessory structure that controls hair growth. It is formed during the embryonic period and is mainly composed of hair papilla, hair matrix, inner root sheath, outer root sheath, bulge area, and hair shaft. The signal of hair follicle morphogenesis originates from the dermis, and then a series of signaling molecules shuttle between epithelial cells and dermal cells, mediate the interaction between these two cell populations, and promote the proliferation, differentiation, and migration of epithelial cells and dermal cells And a series of activities connected with each other, finally forming a complete hair follicle. The growth of hair is a cyclical process, which is divided into growth phase, regression phase and resting phase. The on...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/12C12N15/85
Inventor 邓学梅崔凯杨祖张媛媛葛雅琼
Owner CHINA AGRI UNIV