Scylla paramamosain peroxy gene Sp-PX, amino acid sequence encoded by using scylla paramamosain peroxy gene Sp-PX and clone method

A technology of Scylla peroxygen and Scylla simulans, which is applied in the field of molecular biology and can solve problems such as economic losses in the breeding industry

Inactive Publication Date: 2014-03-26
INNER MONGOLIA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the past 20 years, the breeding process of blue crabs has been plagued by white spot virus, Vibrio and other bacteria. The infection of blue crabs by bacteria and viruses has brought great economic losses to the related farming industry
However, unlike higher vertebrates, blue crabs can be vaccinated to defend against the invasion of bacteria and viruses, and the scale of crab disease can only be alleviated by finding measures that can enhance the antibacterial and antiviral capabilities of blue crabs.

Method used

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  • Scylla paramamosain peroxy gene Sp-PX, amino acid sequence encoded by using scylla paramamosain peroxy gene Sp-PX and clone method
  • Scylla paramamosain peroxy gene Sp-PX, amino acid sequence encoded by using scylla paramamosain peroxy gene Sp-PX and clone method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Cloning of the peroxin gene Sp-PX in the hepatopancreas tissue of Scylla pseudocaveus

[0023] (1) Select the hepatopancreas tissue of Scylla pseudocavena as the experimental material, and use the Total RNA Extractor (Trizol) animal total RNA rapid extraction kit provided by Shanghai Sangon Bioengineering Co., Ltd. to extract total RNA to obtain the Scylla pseudocavena Total RNA samples from hepatopancreas tissue;

[0024] (2) Using the total RNA sample of the hepatopancreas tissue of Scylla pseudocavena obtained in step (1) as a template, use the one-step RT-PCR amplification kit provided by Shanghai Sangon Bioengineering Co., Ltd. to perform reverse transcription synthesis of cDNA , to obtain the cDNA sample of the hepatopancreas tissue of Scylla pseudocaveus;

[0025] (3) Design the front and rear primers Sp-PX-F and Sp-PX-R for amplifying the Sp-PX coding region of the peroxin gene of Scylla pseudomae:

[0026] Sp-PX-F: 5′-TACTCA GAATTC ATGAAGGGGCTACTGT...

Embodiment 2

[0031] Example 2 Cloning of the histoperoxygen gene Sp-PX in the gills of the mud crab

[0032] (1) Select the gill tissue of the mud crab as the experimental material, and use the Total RNA Extractor (Trizol) animal total RNA rapid extraction kit provided by Shanghai Sangong Bioengineering Co., Ltd. to extract the total RNA to obtain the gills of the mud crab. Tissue total RNA samples;

[0033] (2) Using the total RNA sample of the gill tissue of Scylla pseudocavena obtained in step (1) as a template, use the one-step RT-PCR amplification kit provided by Shanghai Sangon Bioengineering Co., Ltd. to carry out reverse transcription synthesis of cDNA, Obtain the cDNA sample of the gill tissue of the mud crab;

[0034] (3) Design the front and rear primers Sp-PX-F and Sp-PX-R for amplifying the Sp-PX coding region of the peroxin gene of Scylla pseudomae:

[0035] Sp-PX-F: 5′-TACTCA GAATTC ATGAAGGGGCTACTGTTCC-3′

[0036] Sp-PX-R: 5′-TACTCA CTCGAG CTAGTACTTCTTTGGTCCCCCAA-3′

...

Embodiment 3

[0040] Example 3 Cloning of the eye stalk histoperoxide gene Sp-PX of Scylla pseudocaveus

[0041] (1) The eye stalk tissue of Scylla pseudocavena was selected as the experimental material, and total RNA was extracted using the Total RNA Extractor (Trizol) animal total RNA rapid extraction kit provided by Shanghai Sangon Bioengineering Co., Ltd. to obtain Scylla pseudocarpus Total RNA samples of crab eye stalk tissue;

[0042] (2) Using the total RNA sample of the eye stalk tissue of Scylla pseudocavena obtained in step (1) as a template, use the one-step RT-PCR amplification kit provided by Shanghai Sangon Bioengineering Co., Ltd. to perform reverse transcription synthesis of cDNA , to obtain the cDNA sample of the eye stalk tissue of Scylla pseudocaveus;

[0043] (3) Design the front and rear primers Sp-PX-F and Sp-PX-R for amplifying the Sp-PX coding region of the peroxin gene of Scylla pseudomae:

[0044] Sp-PX-F: 5′-TACTCA GAATTC ATGAAGGGGCTACTGTTCC-3′

[0045] Sp-PX-...

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Abstract

The invention relates to a scylla paramamosain peroxy gene Sp-PX, an amino acid sequence encoded by using the scylla paramamosain peroxy gene Sp-PX and a clone method. The overall length of a cDNA sequence of the gene is obtained by primer design, PCR (Polymerase Chain Reaction) amplification reaction and company sequence measurement, the nucleotide sequence of the scylla paramamosain peroxy gene Sp-PX has 2331bp, and the amino acid sequence encoded by using the scylla paramamosain peroxy gene Sp-PX has 776 amino acid residues. A research on the scylla paramamosain peroxy gene Sp-PX has important significance for further perfecting the theoretical basis of innate immune systems of invertebrates and can also provide theoretical foundation for finding immunization strategy of reinforcing the antibacterial ability and anti-virus capability of the scylla paramamosain.

Description

technical field [0001] The invention relates to a mud crab peroxin gene Sp-PX, its coded amino acid sequence and a cloning method, and belongs to the technical field of molecular biology. Background technique [0002] Invertebrates are a class of animals with a relatively low evolutionary status. The significant difference from vertebrates is that they do not have a spine. In fact, it is the emergence of the spine that led to the renewal of the immune system in the history of biological evolution. The original innate immunity that relied on genetic inheritance has evolved into acquired immunity that can be continuously enriched. The emergence of acquired immunity has enabled organisms to have a wider range of immune forms in the evolution process, and at the same time transformed from a passive defense type of immunity to an active acquisition type of immunity. For invertebrates, the innate immune system is the only form of immune defense against the invasion of pathogens s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/53C12N9/02C12N15/10
Inventor 杜志强李新苍任乾
Owner INNER MONGOLIA UNIV OF SCI & TECH
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