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Promoter of bovine alpha-actin gene and application thereof

A promoter and gene technology, applied in the field of genetic engineering, can solve the problem of low transcription activity, achieve the effects of strong promoter activity, increase muscle yield, and improve expression activity

Inactive Publication Date: 2015-05-27
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But its transcriptional activity is still low compared to some strong promoters

Method used

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  • Promoter of bovine alpha-actin gene and application thereof
  • Promoter of bovine alpha-actin gene and application thereof
  • Promoter of bovine alpha-actin gene and application thereof

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Embodiment 1

[0019] 1. Cloning of 170bp ① nucleic acid fragment inside the 262 promoter fragment of bovine α-actin gene

[0020] Using the positive recombinant plasmid pGL3-α-actin262 that has been constructed in our laboratory as a template, use the designed primers (Table 1) to target between -224bp and -55bp upstream of the transcription site in the 262 promoter fragment of the bovine α-actin gene The sequence containing multiple SRE regulatory elements was amplified by PCR, and a 170bp nucleic acid sequence was obtained, and this sequence was named: α-actin170①. Among them, α-actin170① contains KpnI and BamHI double enzyme cutting sites. The PCR reaction program was: pre-denaturation at 94°C for 5 min, denaturation at 94°C for 30 s, annealing for 30 s, extension at 72°C, a total of 35 cycles; final extension at 72°C for 10 min, and cooling to 4°C for storage.

[0021] Table 1 Primers for PCR amplification of α-actin170①

[0022]

[0023] The α-actin170①PCR product was recovered an...

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Abstract

The invention discloses a promoter of a bovine alpha-actin gene and an application thereof, belonging to the technical field of genetic engineering. The promoter is characterized in that the nucleic acid sequence of 170bp containing three SRE positive regulation elements inside the 262bp promoter segment of the bovine alpha-actin gene is subjected to PCR amplification by use of different primers to obtain two nucleic acid sequences of 170bp, and the two nucleic acid sequences of 170bp are connected in series with the upstream of the 262bp promoter segment (namely, 170bp-170bp-262bp) to build a promoter segment of alpha-actin, wherein the nucleotide sequence is shown by Seq ID No:1. The promoter has relatively high promoting activity and skeletal muscle specificity and can remarkably improve the expression activity of the alpha-actin gene in bovine skeletal muscle satellite cell and can be used as an efficient and specific skeletal muscle promoter and applied to the transgenic beef production for increasing the muscle output.

Description

technical field [0001] The invention relates to a promoter of bovine α-actin gene and its application, belonging to the technical field of genetic engineering. Background technique [0002] The regulation of gene expression in eukaryotes is a complex and rigorous process. In the study of transgenic animals, it is of great significance to obtain highly efficient and specific promoters that can be widely used for the expression of foreign genes. A promoter is an integral part of a gene (gene), which controls the initiation time and degree of gene expression (transcription). Promoters are like "switches" that determine the activity of genes. Since the gene is a sequence of nucleotides (nucleotides), the promoter should also be composed of DNA. The promoter itself does not control gene activity, but controls gene activity by binding to such proteins (proteins) called transcription factors. Transcription factors are like a "flag" directing the activity of enzymes (RNA polymer...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/85
Inventor 严云勤佟慧丽李树峰胡倩赵丹丹李光鹏张伟伟
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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