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Gene sequence of directivity and differentiation factor 1 fat cell of pigs

A differentiation factor, gene sequence technology, applied in the field of protein coding sequence, can solve the problem of undiscovered ADD1 gene sequence polymorphism and other problems

Inactive Publication Date: 2006-10-11
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, polymorphisms in the ADD1 gene sequence have not been found so far, and linkage analysis of genotypes and traits at mutation sites and marker-assisted selection cannot be performed.

Method used

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  • Gene sequence of directivity and differentiation factor 1 fat cell of pigs
  • Gene sequence of directivity and differentiation factor 1 fat cell of pigs
  • Gene sequence of directivity and differentiation factor 1 fat cell of pigs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0103] Cloning of porcine ADD1 gene

[0104] 1. Isolation

[0105] The fresh spleen tissue of pigs collected from the slaughterhouse (the breed of "Meishan pig") was quickly frozen in liquid nitrogen, and brought back to the laboratory, stored at -70°C, ready to extract total RNA from the tissue.

[0106] 2. Total RNA isolation (Total RNA isolation)

[0107] (1) Preparation for RNA extraction

[0108] Glass products were soaked in 0.1M NaOH overnight, rinsed repeatedly with tap water, rinsed twice with distilled water, and baked at 180°C for 4 hours.

[0109] Soak the homogenizer and electrophoresis tank with 3% hydrogen peroxide for 20-30 minutes, and then rinse with 0.1% DEPC water. Since non-inactivated DEPC has an impact on the PCR reaction, it can be treated with 0.5% SDS.

[0110] Soak the Tip and EP tube in 0.1% DEPC water overnight, and sterilize by autoclaving to inactivate DEPC.

[0111] The double-distilled water and solution were treated with DEPC, that is, 0....

Embodiment 2

[0128] Sequence analysis and homology comparison of porcine ADD1 gene

[0129] 1. Sequence analysis of porcine ADD1 gene

[0130]After translating the CDS part of the cDNA sequence into amino acid sequence with primer primer 5.0 software, the structure prediction was performed with rpsBlast software on http: / / www.ncbi.nlm.nih.gov / Structure / cdd / wrpsb.cgi. It was found that there is a core structure of helix-loop-helix (Helix-Loop-Helix, HLH) at 320-380 of the amino acid sequence (see attached Figure 4 ).

[0131] 2. Homology analysis of porcine ADD1 gene sequence

[0132] The amino acid sequence and base sequence of the gene were analyzed by Blastn and Blastp software on http: / / www.ncbi.nlm.nih.gov / BLAST / BLAST / Blast.cgi, respectively. As a result, it was found that the base sequence and amino acid sequence of the gene were highly homologous to human and mouse ADD1 genes.

[0133] 3. Construction of phylogenetic tree

[0134] The sequence of ADD1 gene and SREBP2 gene (anot...

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Abstract

A gene sequence of pig adipocyte orientation and differentiation factor 1 used in the technical field of genetic engineering, the ADD1 gene amplified from the total RNA of spleen cells of pigs, its cDNA sequence and the amino acid sequence of the encoded protein, the full length of the gene 3830bp, open reading frame 11bp-3453bp segment, encoding a protein with 1151 amino acids, the non-coding sequence at the 3' end is 10bp long, the non-coding sequence at the 5' end is 366bp long, and there is a helix-loop at 320-380 of the protein sequence - Spiral core structure. In one aspect of the present invention, the protein of the gene can be prepared to study the function of the gene in the process of pig fat cell differentiation and regulation of cholesterol level in the body. On the other hand, it can play a guiding role in the production and breeding of lean-meat pigs and low-cholesterol pigs through the method of marker-assisted selection (MAS).

Description

technical field [0001] The present invention relates to a protein coding sequence used in the technical field of genetic engineering, in particular to a gene sequence of porcine adipocyte orientation and differentiation factor 1. technical background [0002] Fat traits are one of the main indicators of animal growth and meat quality traits, and are related to many economic traits in animal production such as lean meat percentage, growth rate, feed conversion rate, and intramuscular fat. In the long-term production practice, it has long been found that the prices of animal products with different fat contents vary greatly. People's consumption habits have shifted from eating fatty meat to eating lean meat, which makes the economic benefits of obese individuals decline in the process of animal production. Fat has become a by-product of animal production. It reduces the feed conversion ratio and at the same time reduces the economics of the animal production process. Fatty ...

Claims

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Application Information

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IPC IPC(8): C12N15/12C07K14/47C12N15/11
Inventor 潘玉春李长龙孟和赵威
Owner SHANGHAI JIAO TONG UNIV
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