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Nucleotide sequence for coding Bactrian camel cytochrome P450, Bactrian camel cytochrome P450 and application thereof

A technology of nucleotide sequence and cytochrome, applied in the application field of nucleotide sequence, can solve the problems of Bactrian camel's detoxification ability and body temperature regulation, etc.

Inactive Publication Date: 2014-12-03
XINJIANG WANGYUAN CAMEL MILK IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Compared with other animals, camels have a stronger ability to regulate body temperature and can adapt to the harsh environment in the desert, which is related to many factors. Although there are many studies on cytochrome P450 at home and abroad, the special detoxification ability of Bactrian camels and thermoregulation have been poorly studied

Method used

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  • Nucleotide sequence for coding Bactrian camel cytochrome P450, Bactrian camel cytochrome P450 and application thereof
  • Nucleotide sequence for coding Bactrian camel cytochrome P450, Bactrian camel cytochrome P450 and application thereof
  • Nucleotide sequence for coding Bactrian camel cytochrome P450, Bactrian camel cytochrome P450 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Cloning of Bactrian camel CYP450 gene cDNA sequence

[0030] 1. Isolation

[0031] Bactrian camels were collected from Alxa League in Inner Mongolia, slaughtered quickly and liver samples were taken. The tissue samples were quickly frozen in liquid nitrogen and stored at -70°C. They were taken back to the laboratory for extraction of tissue total RNA.

[0032] 2. Total RNA isolation

[0033] (1) Preparation for RNA extraction

[0034] 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.

[0035] Soak the homogenizer and electrophoresis tank with 3% hydrogen peroxide for 20-30 minutes, and then rinse with 0.1% DEPC (diethylpyrocarbonate) water. Since the uninactivated DEPC has an impact on the PCR reaction, 0.5% SDS can be used (sodium lauryl sulfate) treatment.

[0036] Tips and EP tubes were soaked in 0.1% DEPC water overnight, and sterilized by autoclaving...

Embodiment 2

[0057] Example 2: Homology comparison and phylogenetic tree construction of Bactrian camel CYP450 gene coding sequence

[0058] 1. Sequence information and bioinformatics analysis of Bactrian camel CYP450

[0059] The length of the new Bactrian camel olfactory receptor CDS of the present invention is 1591bp, and the detailed sequence is shown in SEQ ID NO: 1, wherein the 29-31 position is the start codon ATG, the 1578-1580 position is the stop codon TGA, and the 29-1580 position is the coded protein domain (CDS). The amino acid sequence of the Bactrian camel olfactory receptor was deduced according to the full-length CDS, with a total of 516 amino acid residues, see SEQ ID NO:2 for details, online prediction (http: / / www.expasy.ch / tools / pi_tool.html) results It shows that its molecular weight is 31738.41 Daltons, and its isoelectric point (pI) is 8.36.

[0060] 2. Construction of CYP450 gene coding sequence phylogenetic tree

[0061] Search on GenBank and obtain the coding p...

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Abstract

The invention discloses a nucleotide sequence for coding a Bactrian camel cytochrome P450, a Bactrian camel cytochrome P450 and application thereof, relating to the technical field of gene engineering. The main technical scheme is as follows: the nucleotide sequence is 29th-1580th sites in SEQ ID NO:1 in the sequence table; or the nucleotide sequence is hybridized with the 29th-1580th nucleotide sequence in SEQ ID NO:1 in the sequence table under highly strict conditions and used for coding the Bactrian camel cytochrome P450; or the nucleotide sequence has more than 90% of homology with the 29th-1580th nucleotide sequence in SEQ ID NO:1 in the sequence table and used for coding the Bactrian camel cytochrome P450. The Bactrian camel cytochrome P450 is coated by the nucleotide sequence.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a nucleotide sequence encoding Bactrian camel cytochrome P450, the encoded Bactrian camel cytochrome P450, and the application of the nucleotide sequence. Background technique [0002] Cytochrome P450 is (cytochromeP450, CYP450) a kind of heme-containing single-chain protein that combines with CO in a reduced state and has an absorption peak at a wavelength of 450nm. It is a superfamily of heme-thiolate proteins. It is mainly found in the microsomes and mitochondria of mammals. It has been found that almost 1000 CYPs are widely distributed in various biological organisms, and about 50 are physiologically functional, which are divided into 17 families and many subfamilies according to the unity of amino acid sequence. It is involved in the metabolism of endogenous substances and exogenous substances including drugs, environmental compounds. In recent years, great pro...

Claims

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Application Information

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IPC IPC(8): C12N15/53C12N9/02C12N15/63C12N15/10
Inventor 陈钢粮
Owner XINJIANG WANGYUAN CAMEL MILK IND
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