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Construction method for DCN gene overexpression vector of cashmere goat

A technology of gene overexpression and construction method, applied in the field of construction of DCN gene overexpression vector

Inactive Publication Date: 2015-11-11
INNER MONGOLIA AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no method for constructing the DCN gene overexpression vector of cashmere goats

Method used

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  • Construction method for DCN gene overexpression vector of cashmere goat
  • Construction method for DCN gene overexpression vector of cashmere goat
  • Construction method for DCN gene overexpression vector of cashmere goat

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Experimental program
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Embodiment Construction

[0014] The technical solutions of the present invention will be further described in detail below in conjunction with specific embodiments.

[0015] 1 Materials and methods

[0016] 1.1 Materials: Cashmere goat skin samples were collected from adult ewes in Inner Mongolia white cashmere goat breeding farm.

[0017] 1.2 Method

[0018] 1.2.1 DCN gene cloning

[0019] Extraction of total RNA from cashmere goat skin, synthesis of the first strand of cDNA, design of primers (DCN-F, DCN-R), using cDNA as a template, using primers DCN-F, DCN-R and Phusion high-fidelity DNA polymerase, according to Conventional methods for PCR amplification. The reaction conditions are: 98°C for 5min; 30 cycles of 98°C for 30s, 59°C for 2min, and 72°C for 2min; 72°C for 10min. PCR amplification products were identified by agarose gel electrophoresis. The nucleotide sequence of DCN-F is shown in SEQ ID NO: 1, and the nucleotide sequence of DCN-R is shown in SEQ ID NO: 2.

[0020] 1.2.2 Construct...

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Abstract

The invention discloses a construction method for a DCN gene overexpression vector of a cashmere goat. The method comprises the following steps: (1) cloning a DCN gene; (2) constructing the overexpression vector pEGFP-Cl / DCN. According to the method, on the basis of cloning a DCN gene cDNA sequence of the cashmere goat, the overexpression vector pEGFP-Cl / DCN is constructed, and foundation is laid for follow-up research of DCN gene functions of the cashmere goat.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for constructing a cashmere goat DCN gene overexpression vector. Background technique [0002] Decorin (decorin, DCN) is a kind of proteoglycan (proteoglycan, PG), which is a kind of pleiotropic molecule. It plays an important role in regulating cell adhesion, migration, proliferation, collagen fiber formation and regulating TGF-β activity. At present, there is no method for constructing a cashmere goat DCN gene overexpression vector. Contents of the invention [0003] The purpose of the present invention is to overcome the defects in the prior art and provide a method for constructing a cashmere goat DCN gene overexpression vector. In this method, the overexpression vector pEGFP-CI / DCN was constructed on the basis of cloning the cDNA sequence of the cashmere goat DCN gene, which laid the foundation for the subsequent study of the function of the cashmere goat DCN gene. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/79C12N15/66
Inventor 王乐乐张燕军王瑞军苏蕊李金泉
Owner INNER MONGOLIA AGRICULTURAL UNIVERSITY
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