Goat TNNT3 (human fast skeletal troponin T) gene and method for cloning complete sequence of gene coding region of goat TNNT3 gene

A cloning method and coding region technology, applied in the field of goat TNNT3 gene and cloning the complete sequence of goat TNNT3 gene coding region, can solve the problem of no goat TNNT3 gene

Inactive Publication Date: 2011-09-14
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] There is no related report about goat TNNT3 gene

Method used

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  • Goat TNNT3 (human fast skeletal troponin T) gene and method for cloning complete sequence of gene coding region of goat TNNT3 gene
  • Goat TNNT3 (human fast skeletal troponin T) gene and method for cloning complete sequence of gene coding region of goat TNNT3 gene
  • Goat TNNT3 (human fast skeletal troponin T) gene and method for cloning complete sequence of gene coding region of goat TNNT3 gene

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Embodiment Construction

[0021] The present invention will be described in detail below in conjunction with specific embodiments.

[0022] Step 1 extracts the total RNA (ribonucleic acid) in the goat leg muscle tissue, and then reverse-transcribes the total RNA into cDNA;

[0023] 1. Collection of Tissue

[0024] Immediately after bleeding from the carotid artery of one-year-old Tianfu mutton sheep, about 1 g of leg muscle was taken and placed in a liquid nitrogen tank for total RNA extraction

[0025] 2. RNA preparation and cDNA synthesis

[0026] Grind the goat leg muscle into powder with liquid nitrogen in the laboratory, put it into a 1.5ml EP tube, and put it in a minus 80 ultra-low temperature refrigerator for later use.

[0027] Take a new 1.5ml EP tube, add 1.2ml of Trizol solution, and then add about 80mg of leg muscle tissue powder. Immediately shake and mix by hand, shake with a shaker for about 30 seconds, and then place at room temperature for 4 minutes.

[0028] Centrifuge at 13500rp...

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Abstract

The invention discloses a goat TNNT3 (human fast skeletal troponin T) gene, and the nucleotide sequence of the goat TNNT3 gene is SEQ (sequence) ID (identity) NO: 1. The sequence of a coding region is SEQ ID No: 2. Amino acid coded by the sequence of the coding region is SEQ ID No: 3. The invention further discloses a method for cloning the goat TNNT3 gene, which comprises the following steps: step 1) extracting total RNA (ribonucleic acid) in goat leg muscle tissues, and then performing reverse transcription on the total RNA to form cDNA (complementary deoxyribonucleic acid); step 2) designing a primer and performing PCR (polymerase chain reaction) reaction; step 3) recovering and purifying a PCR product; and step 4) cloning. By adopting the method, the complete sequence of the coding region of the goat TNNT3 gene can be simply and quickly obtained, thereby filling in the blank of the gene on a goat, and laying a foundation for further researching the gene of the goat.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a goat TNNT3 gene and a method for cloning the complete sequence of the goat TNNT3 gene coding region. Background technique [0002] Fast-twitch troponin (TNNT3) is a member of the calcium-binding protein complex present in skeletal muscle. Troponin T (TNT), together with troponin C and troponin I, forms a ternary troponin complex that plays a key role in muscle contraction and relaxation. TNNT3 is an asymmetric protein with a globular C-terminal domain, it has a binding site for tropomyosin, and is the protein responsible for binding the troponin complex to tropomyosin. Coordinates the calcium ion concentration-dependent ATPase activity of myofilaments, thereby playing a role in muscle cell contraction and relaxation. [0003] There is no related report about goat TNNT3 gene. Contents of the invention [0004] The technical problem to be solved by the present invention is to pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/10C07K14/47
Inventor 陈浩林徐刚毅汪代华吴婷婷
Owner SICHUAN AGRI UNIV
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