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Chinese prawn heat shock protein 70 gene and its clone method

A technology of heat shock protein and Chinese prawn, applied in the field of molecular biology, can solve the problem that the output of shrimp cannot meet the large demand of seafood, and achieve the effect of improving disease resistance and anti-adversity ability, improving adaptability and promoting development.

Inactive Publication Date: 2004-10-06
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In this case, the output of shrimp is far from meeting people's large demand for seafood.

Method used

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  • Chinese prawn heat shock protein 70 gene and its clone method

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Effect test

Embodiment 1

[0020] A cloned Chinese prawn heat shock protein (HSP) 70 gene has the sequence shown in SEQ ID NO 1:

[0021] Homologous gene one:

[0022] (1) Information of SEQ ID NO.1 (see sequence listing)

[0023] (a) Sequence features:

[0024] * Length: 2090 base pairs

[0025] * Type: nucleic acid

[0026] * Chain type: double chain

[0027] * Topology: Linear

[0028] (b) Molecular type: cDNA

[0029] (c) Assumption: No

[0030] (d) Antisense: no

[0031] (e) Original source: Chinese prawns (Fenneropenaeus chinensis)

[0032] (2) Information of SEQ ID NO.2 (see sequence listing)

[0033] (a) Sequential features

[0034] * Length: 652 amino acids

[0035] * Type: amino acid

[0036] * Chain type: single chain

[0037] * Topology: Linear

[0038] (b) Molecule type: peptide

Embodiment 2

[0039] Example 2 Cloning method of Chinese prawn HSP70 gene

[0040] The method for cloning the above-mentioned HSP70 gene from Penaeus chinensis of the present invention comprises:

[0041] (1) Total RNA was extracted from the liver tissue of Penaeus chinensis;

[0042] (2) Then carry out cDNA first-strand synthesis;

[0043] (3) Using a pair of primers to perform PCR to obtain the shrimp HSP70 gene.

[0044] in:

[0045] The forward primer P1: 5'>GAG AGA AGA GCA GAC GTG TTA CA<3';

[0046] Reverse primer P4: 5′>TGA TCT ACG ATG GGT CGA TIT G<3′

[0047] In order to obtain high-quality RNA, the present invention adopts the following RNA extraction procedures, and the specific operating conditions are as follows:

[0048]1. Total RNA extraction: 1) Preparation of extract: RNA extract contains 4M guanidine isothiocyanate, 0.5% sodium lauryl sarcosine, 26mM sodium citrate (PH4.0), 0.1M β-mercapto Ethanol; specific preparation method: take by weighing 94.6 grams of guanidine...

Embodiment 3

[0072] The difference from Example 2 is:

[0073] Total RNA was extracted from the eye stalk tissue of Penaeus chinensis.

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Abstract

A Chinese prawn heat shock protein gene 70 is disclosed, which has the SEQ ID No.1 base sequence and SEQ ID No.2 amino acid sequence. Its cloning process includes extracting general RNA from tissue of Chinese prawn, synthesizing the first chain of cDNA, and using a pair of primer for RT-PCR.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, in particular to a Chinese prawn heat shock protein gene 70 and a cloning method thereof. Background technique [0002] Heat shock protein (HSP) is a group of proteins with highly conserved sequences encoded and synthesized by heat shock genes in all cells under stress. They can make cells resist more severe stress damage. Among them, HSP70 is the most conserved and important heat shock protein family. As a molecular chaperone, HSP70 participates in the correct folding and assembly of new cell proteins, the unfolding of misfolded proteins, and the renaturation or clearance of denatured proteins. Heat shock protein is also expressed at a certain level under normal physiological conditions, and its expression is up-regulated under the stimulation of factors such as heat shock, heavy metal ions, virus or bacteria infection, and participates in the protection of body cells. It is a non-spe...

Claims

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Application Information

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IPC IPC(8): C07H21/04C12N15/12C12P19/34
Inventor 相建海焦传珍王在照李富花
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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