Lateolabrax japonicus actin gene sequence

A technology of muscle and protein, applied in the field of gene cloning in molecular biology

Inactive Publication Date: 2006-12-27
SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there are few studies on the genes of perch cell

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0027] 1. Extraction of total RNA

[0028] Fresh and healthy perch (about 400 g in weight) was taken for 2 days in the laboratory (water temperature was about 24°C, air pump was inflated), and 200 μL of lipopolysaccharide (LPS, 10 μg / mL) was injected from the chest cavity. After 6 hours of stimulation, the fish body was dissected to take out about 100 mg of spleen, which was put into 1 mL Trizol (Gibco, Japan) for homogenization, and total RNA was extracted according to the kit instructions.

[0029] 2. Synthesis of the first strand of cDNA

[0030] Take 5 μg of total RNA of perch and mix with 1 μL (10 pmol / L) of reverse transcription primer (oligo-dT linker primer), heat at 70 ° C for 5 min, place it on ice immediately, and then add 5 × buffer, 2.5 mmol / L dNTP mixture , Ribonuclease Inhibitor, M-MLV reverse transcriptase, the reaction system is 25 μL. The reaction process was 60 minutes at 42°C, 15 minutes at 70°C, and finally stored at -80°C for later use.

[0031] 3. Pri...

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PUM

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Abstract

This invention is the expressed sequence of actins' gene, which is the degenerate primer of conserved regions design from CDS sequence of related species actins' gene. It extracts total RNA from spleen of weever, then amplificates by PCR approach and clones to actins of weever. This gene sequence not only lays the foundation for the research of the effect of fish actins during DNA transcription, but also can gain the active recombinant protein so as to provide theory basic of the research of the new type immunity adjuvar.

Description

technical field [0001] The invention relates to the field of gene cloning in molecular biology, in particular to a gene sequence of sea bass actin. Background technique [0002] Actin is defined as a molecular engine that is involved in cell movement, cell division, and wound healing; at the same time, actin is also involved in DNA transcription and can recruit RNA polymerase II to act as a molecular engine when RNA polymerase II begins transcription. If actin is repressed, transcription cannot begin. The process of DNA transcription is central to all cellular activities. These functions will allow us to intervene effectively when genetic transcription goes awry; actin can function as a cofactor with viral enzymes that degrade the cytoskeleton. [0003] At present, with the development of mariculture industry, the problem of disease is becoming more and more serious. Therefore, starting from the fish itself, cloning related functional genes, obtaining purified and active r...

Claims

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Application Information

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IPC IPC(8): C12N15/12C07K14/435
Inventor 邱丽华江世贵张汉华
Owner SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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