Cloning method of schizothorax prenati gene CART

A technology of Schizothorax, CART-1, which is applied in the field of CART gene cloning of Schizothorax, can solve problems such as the lack of research on CART genes

Inactive Publication Date: 2014-09-24
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, research on CART genes in fish is still relatively lacking, and gene cloning has only been carried out in a few fish such as zebrafish, goldfish, and Atlantic salmon.

Method used

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  • Cloning method of schizothorax prenati gene CART
  • Cloning method of schizothorax prenati gene CART
  • Cloning method of schizothorax prenati gene CART

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] The method for cloning Schizothorax CART gene specifically comprises the following steps:

[0019] 1. Extraction of total RNA from brain tissue

[0020] The extraction process of total RNA was carried out strictly according to the requirements of aseptic operation. The specific operation steps are as follows:

[0021] (1) Take out the frozen tissue from the -80°C refrigerator, take about 100 mg, put it into a high-temperature sterilized mortar, add liquid nitrogen, and grind it into powder;

[0022] (2) Add 1mL of pre-cooled RNAisoPlus lysate to a 1.5mL centrifuge tube, quickly place the tissue powder in the centrifuge tube, and shake vigorously for 5 minutes to fully dissolve it;

[0023] (3) Centrifuge at 12000g for 5min at 4°C, carefully absorb the supernatant, and transfer it to a new 1.5mL centrifuge tube.

[0024] (4) Add 0.2 mL of chloroform to the centrifuge tube, close the cap of the centrifuge tube tightly, shake vigorously by hand for 15 s, wait until the ...

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Abstract

The invention relates to the field of biotechnology, and in particular relates to a cloning method of a schizothorax prenati gene CART. Three subtypes of the gene CART are obtained via the cloning, the sequencing and the expression analysis of the schizothorax prenati gene CART. The invention provides a method for efficiently and conveniently obtaining an overall-length sequence of the schizothorax prenati gene CART. The blank of the gene CART on schizothorax prenati is compensated, thereby laying a foundation for further research of the schizothorax prenati gene CART.

Description

technical field [0001] The invention relates to the field of biological technology, in particular to a method for cloning the CART gene of Schizothorax chinensis. Background technique [0002] With the deepening and improvement of aquatic animal nutrition research, in recent years, fish feeding regulation has gradually become a research hotspot in nutrition. So far, different scholars have studied nutrition and animal growth performance, physiological parameters, and biochemical enzymes and A lot of research work has been done on the relationship between metabolism and other aspects, but at the molecular level, especially the research on the relationship between feeding-related genes and fish energy metabolism is still quite lacking. [0003] Cocaine-andamphetamine-regulated transcript peptides (CART) is an endogenous neuropeptide discovered in recent years. Previous studies have suggested that CART is widely distributed in the central nervous system and peripheral tissues,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/63C12N1/21C12N1/19C12N5/10C07K14/46
Inventor 陈德芳袁登越李志琼
Owner SICHUAN AGRI UNIV
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