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Construction and application of a bovine klf3 gene eukaryotic overexpression vector

An overexpression vector, expression vector technology, applied in the field of genetic engineering, can solve the problem of unclear influence rules and so on

Active Publication Date: 2021-11-30
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are a few research reports on KLF3 gene in marker-assisted selection of growth performance of yellow cattle (for example, Qinchuan cattle) and breeding of new elite breeds. Theoretical basis and practical approach for regulating cattle muscle growth and development traits and beef quality

Method used

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  • Construction and application of a bovine klf3 gene eukaryotic overexpression vector
  • Construction and application of a bovine klf3 gene eukaryotic overexpression vector
  • Construction and application of a bovine klf3 gene eukaryotic overexpression vector

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Embodiment Construction

[0034] The present invention will be described in further detail below in conjunction with the accompanying drawings and examples, which are only used to explain the present invention rather than limit the protection scope of the present invention.

[0035](1) Construction of plasmid-type recombinant overexpression vector pcDNA3.1-KLF3

[0036] 1. Materials and methods:

[0037] 1.1. Instrument:

[0038] Ultra-clean bench, biochemical incubator, gene amplification instrument, PTC-200 single-slot gradient gene amplification instrument, Heraeus refrigerated high-speed centrifuge (Germany), Bio-Rad gel imaging analyzer (USA), CO 2 Incubator, HZS-H water bath shaker (Harbin), Eppendorf pipette, DYY-Ⅲ steady voltage and steady current electrophoresis instrument (Beijing Liuyi), DYY-Ⅲ31A and DYY-Ⅲ28D electrophoresis tank (Beijing Liuyi), manufacturing Ice instrument, MDF-382E ultra-low temperature refrigerator (Sanyo, Japan), Eppendorf desktop high-speed centrifuge, Sartorious ele...

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Abstract

The invention discloses the construction and application of a bovine KLF3 gene eukaryotic overexpression vector. Primers were designed for the coding region of Qinchuan cattle KLF3 gene, and the corresponding bovine KLF3 gene was amplified; pcDNA3.1‑KLF3 recombinant overexpression vector was constructed and transfected into cells. By cloning the bovine KLF3 gene and constructing a eukaryotic overexpression vector, the invention can be applied to bovine KLF3 gene function research, identification and regulation of muscle growth and development.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering and relates to a plasmid-type recombinant overexpression vector pcDNA3.1-KLF3 and a construction method thereof. The overexpression vector can be used in functional identification of transformed seed cells and bovine KLF3. Background technique [0002] At present, gene expression has become a mainstream technology in biology, medicine and drug development research. Gene overexpression as "reasonable reversal" of gene knockout. Overexpression of a gene ensures that more messenger RNA is produced, which in turn produces more protein or downstream products. It can positively regulate the performance of a gene, which is convenient for studying the function of a gene in an organism. [0003] At present, many plasmid vectors that can realize gene overexpression have been developed. The pcDNA3.1(+ / -) vector is one of the most commonly used mammalian expression vectors. It utilizes the stro...

Claims

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Application Information

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IPC IPC(8): C12N15/85C12N15/66C12Q1/6888G01N33/68C12Q1/02C12N5/10
CPCC07K14/4703C12N5/0658C12N15/66C12N15/85C12N2510/00C12Q1/6888G01N33/5061G01N33/68G01N2333/4704
Inventor 黄永震徐嘉威王大会刘贤贺花张子敬于翔吕世杰王二耀陈宏雷初朝胡沈荣
Owner NORTHWEST A & F UNIV
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