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Bactrian camel H-FABP protein gene, recombinant protein and cloning method thereof

A technology for recombining proteins and Bactrian camels, which is applied in the field of genetic engineering, can solve problems such as cloning and obtaining the encoded protein sequence of Bactrian camel H-FABP gene.

Inactive Publication Date: 2013-04-03
XINJIANG WANGYUAN CAMEL MILK IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] There are few reports on the genetic variation of the H-FABP gene in animals. So far, no reports have been found on the cloning of the H-FABP gene-encoded protein sequence of the Bactrian camel and the study on the evolution of the H-FABP gene in animals.

Method used

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  • Bactrian camel H-FABP protein gene, recombinant protein and cloning method thereof
  • Bactrian camel H-FABP protein gene, recombinant protein and cloning method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1, a Bactrian camel H-FABP protein gene, the nucleotide sequence of the gene has the sequence shown in SEQ ID NO.1. The new gene sequence obtained by using the total RNA in the liver tissue of Bactrian camel as a template, referring to the homologous sequence of the H-FABP protein gene of bovine, human, mouse and other species to design primers, and performing reverse transcription PCR. The cDNA sequence of camel H-FABP protein gene is shown in SEQ ID NO.1.

[0031] According to actual needs, the above-mentioned Bactrian camel H-FABP protein gene can be further optimized or / and improved:

[0032] The nucleotide sequence of the Bactrian camel H-FABP protein gene has the sequence of 46-661 in SEQ ID NO.1. In SEQ ID NO.1, the length of the RT-PCR product is 708bp, and the electrophoresis results are shown in Figure 1, wherein the 46-48 position is the start codon ATG, the 659-661 position is the stop codon TGA, and the 46-661 position is Coding protein domain (CD...

Embodiment 2

[0033] Example 2, a recombinant protein of Bactrian camel H-FABP protein gene.

[0034] According to actual needs, the recombinant protein of the above-mentioned Bactrian camel H-FABP protein gene can be further optimized or / and improved:

[0035] The amino acid sequence of the recombinant protein has the sequence shown in SEQ ID NO.2. See SEQ ID NO.2 for the protein coding sequence translated from the coding sequence (CDS) in the cDNA sequence of the Bactrian camel H-FABP protein gene according to the common codons.

[0036] The recombinant protein is a polypeptide having the amino acid sequence shown in SEQ ID NO.2, or a conservative variant polypeptide thereof, or an active fragment thereof, or an active derivative thereof.

[0037] By designing a pair of primers to obtain the Bactrian camel myosin gene from the Bactrian camel H-FABP protein, the nucleotide sequence information of the primer pair is as follows:

[0038]Forward primer, SEQ ID NO.3 (forward): 5'-ATGGTGGACGC...

Embodiment 3

[0040] Embodiment 3, a kind of cloning method of Bactrian camel H-FABP protein gene is characterized in that carrying out according to the following steps:

[0041] The first step, total RNA extraction

[0042] 1. Isolation

[0043] 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.

[0044] 2. Total RNA isolation

[0045] (1) Preparation for RNA extraction

[0046] 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.

[0047] Soak the homogenizer and electrophoresis tank with 3% hydrogen peroxide for 20-30 minutes, and then rinse with 0.1% DEPC water. Since the uninactivated DEPC will affect the PCR reaction, it can be treated with 0.5% SDS.

[0048] Tips an...

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Abstract

The invention relates to the technical field of genetic engineering and discloses a bactrian camel H-FABP protein gene, a recombinant protein and a cloning method thereof. The H-FABP gene sequence of other species is referred, a cDNA sequence of the bactrian camel H-FABP protein gene is obtained through clone sequencing, the CDS sequence of the H-FABP gene of various species is subjected to homologous comparison, the structural features of the H-FABP gene are known and verified, and basic data are provided for a relationship for researching the H-FABP gene and the bactrian camel lipid metabolism. When H-FABP is combined with the traditional markers such as cTnI and CK-MB for detection, the H-FABP is detected in the early stage, and the cTnI is detected in the restoration stage. A patient suffering from cardiac event high risk is screened, the positive treatment strategy is conveniently taken, the expenditure of the patient and the lab resources can be saved, and reasonable and complementary monitoring of H-FABP and cTnI is an ideal choice of clinical treatment of ischemic heart disease.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, and relates to a Bactrian camel H-FABP protein gene, a recombinant protein and a cloning method thereof. Background technique [0002] Heart fatty acid-binding protein (heart fatty acid-binding protein, H-FABP), also known as FABP3, the gene encoding this protein is a type that has been studied more in the FABPs family, mainly expressed in cardiac muscle, skeletal muscle and mammary gland . The coding regions of H-FABP genes in different species have high homology, which is not only reflected in the encoded amino acids, but also in the DNA sequence, the number of exons and introns, and the number of corresponding exons. The size of the corresponding intron is the same, but the size of the corresponding intron varies greatly among species. At present, the H-FABP genes of many species have been cloned and mapped, among which the human H-FABP gene is located on chromosome 1 p32-p33, an...

Claims

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Application Information

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IPC IPC(8): C12N15/12C07K14/47C12N15/10
Inventor 陈钢粮
Owner XINJIANG WANGYUAN CAMEL MILK IND
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