Eriocheir sinensis EsSXL gene, amplification primer group thereof and amplification method

A Chinese mitten crab, primer set technology, applied in the fields of botany equipment and methods, biochemical equipment and methods, genetic engineering, etc., can solve the problem of not being able to express SXL and so on

Inactive Publication Date: 2013-03-27
FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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Problems solved by technology

In the case of fruit flies, SXL Genes are expressed in both sexes, but only in functional form in females, whereas in males SXL When the mRNA precursor is spliced, the third exon is retained, and there is a terminator in the third exon, so that in SXL Causes premature termination during mRNA translation, unable to express complete and active SXL

Method used

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  • Eriocheir sinensis EsSXL gene, amplification primer group thereof and amplification method
  • Eriocheir sinensis EsSXL gene, amplification primer group thereof and amplification method
  • Eriocheir sinensis EsSXL gene, amplification primer group thereof and amplification method

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Embodiment Construction

[0029] Below in conjunction with embodiment, further set forth the present invention.

[0030] Eriocheir sinensis in the present invention is provided by Xuyi River Crab Farm in Jiangsu Province, reagents can be purchased from the market, reagents such as RNA purification are purchased from Promega Company of the United States, etc., 3' RACE and 5' RACE kits are purchased from Bao Biological (Dalian) Co., Ltd. Company (Takara).

[0031] Extraction of total RNA: Take 100 mg of testis tissue of Eriocheir sinensis, grind it in a mortar and add liquid nitrogen, and extract total RNA by Trizol method. Trizol reagent is from Invitrogen Company, and the extraction method is according to the instruction manual.

[0032] Cloning of the full-length cDNA sequence:

[0033] A 511bp SXL gene homologous sequence (GenBank accession number: JR771373.1) was screened from the GenBank Chinese mitten crab nucleic acid database, and this partial sequence was used as an intermediate sequence to de...

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Abstract

The invention discloses an Eriocheir sinensis EsSXL gene, an amplification primer group thereof and an amplification method. A full-length cDNA (complementary Deoxyribose Nucleic Acid) sequence of the EsSXL gene is cloned from an Eriocheir sinensis testis tissue by using the primer group provided by the invention, and the EsSXL gene comprises two variable spliceosomes EsSXL1 and EsSXL2, and lays an important foundation for the research of the important function of the function of the Eriocheir sinensis EsSXL gene in a sex differentiation process of the Eriocheir sinensis EsSXL gene.

Description

technical field [0001] The invention relates to the field of gene cloning, in particular to Chinese mitten crab QUR gene, including two alternative splice forms ESXL1 and EsXL2 , its amplification primer set and amplification method. Background technique [0002] Chinese mitten crab ( Eriocheirsinensis ) belongs to the phylum Arthropoda, Crustacea, Decapod, Reptilia, Brachycephala, Square Crabidae, Bacteridae, Eriocheidae, commonly known as river crabs. River crab is the freshwater crab with the largest aquaculture output in my country. It is widely distributed in the brackish water or fresh water of the southeast coast of China. Its meat is delicious, unique in flavor and rich in nutrition, and is deeply loved by the masses. Since the mass production of crab seedlings was realized in my country in the early 1980s, the production scale has expanded year by year. In 2008, the total production of river crabs reached 518,400 tons, and the direct economic output value exc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/11C12N15/10
Inventor 沈怀舜周鑫柏爱旭任秀芳
Owner FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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