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Pelteobagrus fulvidraco Cyp24a1 gene and overall-length sequence obtaining method thereof

A technology of pelagic catfish and sequence, applied in the field of genetic engineering, can solve problems such as the lack of Cyp24a1 gene research

Inactive Publication Date: 2016-05-04
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research on Cyp24a1 gene in fish is still relatively lacking.

Method used

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  • Pelteobagrus fulvidraco Cyp24a1 gene and overall-length sequence obtaining method thereof
  • Pelteobagrus fulvidraco Cyp24a1 gene and overall-length sequence obtaining method thereof
  • Pelteobagrus fulvidraco Cyp24a1 gene and overall-length sequence obtaining method thereof

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Embodiment Construction

[0097] In order to better explain the present invention, the main content of the present invention is further clarified below in conjunction with specific examples, but the content of the present invention is not limited to the following examples.

[0098] The method for obtaining the full-length sequence of the Cyp24a1 gene of yellow catfish comprises the following steps:

[0099] 1. Extraction of total RNA from liver tissue

[0100] The extraction process of total RNA was carried out in strict accordance with the requirements of aseptic operation, including the following steps:

[0101] (1) Take about 100 mg of liver tissue from the -80°C refrigerator, put it into a mortar that has been sterilized at high temperature, add liquid nitrogen and grind it into powder;

[0102] (2) Add 1mL of pre-cooled RNAisoPlus lysate to a 2mL centrifuge tube, then quickly add the tissue powder into the centrifuge tube, shake vigorously to dissolve it fully, and let it stand at room temperatur...

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Abstract

The invention discloses a pelteobagrus fulvidraco Cyp24a1 gene and an overall-length sequence obtaining method thereof. The overall length of the Cyp24a1 sequence length is 2180 bp, and 506 amino acids are encoded. A molecular formula of pelteobagrus fulvidraco Cyp24a1 protein is C2602H4112N704O740S26, the size of molecular weight is 57.93 kDa, an isoelectric point is 8.82, and the total average hydrophobic index is -0.351. Prediction and analysis on the structure and functions of a pelteobagrus fulvidraco Cyp24a1 protein shows that the Cyp24a1 protein is free of a signal peptide sequence and a transmembrane structure domain. NCBI comparison of pelteobagrus fulvidraco Cyp24a1 protein shows that the homologies with channel catfish and zebra fish are up to 94% and 74% respectively. The expression of the Cyp24a1 gene in the liver is highest, and the expression of the Cyp24a1 gene in the muscles and intestines is traced. The pelteobagrus fulvidraco Cyp24a1 gene utilizes an RACE technology to fill the blank of the gene in pelteobagrus fulvidraco, and a foundation is laid for further research on the gene for fish functions.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a Cyp24a1 gene of yellow catfish and a method for obtaining a full-length sequence thereof. Background technique [0002] Vitamin D belongs to sterol derivatives, has anti-rickets effect, and is an essential vitamin for animals. Fish cannot synthesize vitamin D and must be supplied by food. Due to fish's response to vitamin D 3 Utilization rate is higher than that of vitamin D 2 , so vitamin D 3 It is the main storage form of vitamin D in fish. Vitamin D 3 After being absorbed by the intestines of fish, it is transported to the liver by chylomicrons through the blood. In the liver, vitamin D 3 Hydroxylation at position 25 to form 25-hydroxyvitamin D 3 [25(OH)D 3 ]. 25(OH)D 3 25-Hydroxyvitamin D 3 -1α-Hydroxylase (CYP24A1) catalyzes conversion to biologically active 1α,25-dihydroxyvitamin D 3 [1α,25(OH) 2 D. 3 ]. [0003] Several in vivo studies have shown that v...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/02C12N15/53C12N15/10
CPCC12N9/0073C12N15/1096C12Y114/13013
Inventor 王春芳陈沛何绪刚谢从新李大鹏
Owner HUAZHONG AGRI UNIV