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Freshwater shrimp KSPI (Kazal-type Serine Proteinase Inhibitor) gene, and amplification method and amplification primer group thereof

A primer set, a technology for green shrimp, applied in genetic engineering, plant genetic improvement, botanical equipment and methods, etc.

Inactive Publication Date: 2012-03-07
FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the molecular research of freshwater shrimp mostly focuses on the genetic diversity of the population, and there are few studies on its gene cloning, and the research on the KSPI gene clone of freshwater shrimp has not been reported yet.

Method used

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  • Freshwater shrimp KSPI (Kazal-type Serine Proteinase Inhibitor) gene, and amplification method and amplification primer group thereof

Examples

Experimental program
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Embodiment 1

[0055] Extraction of total RNA: Take the testis tissues of freshwater shrimp one by one, quickly put them into a pre-cooled mortar filled with liquid nitrogen, take a total of about 30 mg of testicular tissues from three freshwater shrimps, dissect quickly and pay attention to adding liquid nitrogen; total RNA The extraction was carried out using the RNAiso Plus extraction reagent from Takara Company combined with the traditional phenolic extraction method, and the extraction method was referred to the instruction manual.

[0056] cDNA first-strand synthesis: according to the instructions of Shanghai Sangon Bioengineering Co., Ltd. (Sangon) cDNA synthesis kit.

[0057] Cloning of the full-length cDNA sequence of the freshwater shrimp KSPI gene:

[0058] Primers were designed according to the KSPI gene fragment screened in the cDNA library of the testis of the freshwater shrimp constructed in our laboratory, and were used for rapid amplification of the 3' end and the 5' end to ...

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Abstract

The invention relates to the field of gene cloning, and particularly relates to a freshwater shrimp KSPI (Kazal-type Serine Proteinase Inhibitor) gene, and an amplification method and amplification primer group thereof. By utilizing the primer group provided by the invention, a full-length cDNA sequence of the KSPI gene is cloned from a freshwater shrimp spermary tissue, thereby laying a foundation for the research on the function of the freshwater shrimp KSPI gene, and simultaneously accumulating background data for the research on the procreation of shellfish.

Description

technical field [0001] The invention relates to the field of gene cloning, in particular to the green shrimp KSPI gene, its amplification method and amplification primer set. [0002] Background technique [0003] Serine proteinase inhibitor (SPI) is a kind of protein widely present in various organisms, and plays an important role in metamorphosis, wound healing, innate immunity and other processes. Serine protease inhibitors mainly include serpin, α-macroglobulin, Kazal-type, Kunitz-type and other inhibitor families, among which Kazal-type serine proteinase inhibitor (KSPI) is the most widely studied family of enzyme inhibitors one. KSPI was first discovered in mammals, and then in Procambarus clarkii ( Procambarus clarkii ),Vannamei( Litopenaeus vannamei ), Penaeus monodon ( Penaeus monodon ), Macrobrachium rosenbergii ( Macrobrachium rosenbergii ), Toxoplasma gondii ( Toxoplasma gondii ) and silkworm ( Bombyx mori ) and other animals have been identified. K...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/11C12N15/10
Inventor 傅洪拓张响乔慧蒋速飞
Owner FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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