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Freshwater shrimp MnSG (Macrobrachium nipponense Sperm Gelatinase) gene, and amplification method and amplification primer group thereof

A primer set, green shrimp technology, applied in genetic engineering, plant genetic improvement, botanical equipment and methods, etc., can solve the problems of few gene cloning research, and the research on green shrimp sperm protease-related gene cloning has not yet been reported.

Inactive Publication Date: 2012-03-07
FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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Problems solved by technology

At present, most of the molecular studies of freshwater shrimp focus on the genetic diversity of the population, and there are few studies on its gene cloning, and the research on the cloning of the sperm protease-related genes of freshwater shrimp has not been reported yet.

Method used

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  • Freshwater shrimp MnSG (Macrobrachium nipponense Sperm Gelatinase) gene, and amplification method and amplification primer group thereof

Examples

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Embodiment 1

[0053] 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.

[0054]cDNA first-strand synthesis: according to the instructions of Shanghai Sangon Bioengineering Co., Ltd. (Sangon) cDNA synthesis kit.

[0055] Cloning of the full-length cDNA sequence of the freshwater shrimp MnSG gene:

[0056] Design primers according to the MnSG gene fragments screened in the cDNA library of the testis of the freshwater shrimp constructed in our laboratory, for rapid amplification of the 3' end and the 5' end, and obtain the full-len...

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Abstract

The invention relates to the field of gene cloning, and particularly relates to a freshwater shrimp MnSG (Macrobrachium nipponense Sperm Gelatinase) 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 MnSG gene is cloned from a freshwater shrimp spermary tissue, thereby laying a foundation for the research on the function of the freshwater shrimp MnSG 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 MnSG gene, its amplification method and amplification primer set. [0002] Background technique [0003] In eukaryotes, fertilization refers to the mutual fusion of reproductive cells of both sexes. Fertilization includes a series of complex physiological processes such as sperm capacitation, acrosome reaction, fusion through the zona pellucida, and fusion of egg membranes. The proteases on the surface of sperm play an important regulatory role in these processes. Research on mice by Honda et al. showed that there is a protease with gelatin hydrolysis activity on mouse sperm, and this protease is considered to play an important role in the passage of sperm through the egg membrane. In the study of rabbits, Stambaugh et al. found that a trypsin on the acrosome of rabbit sperm is necessary for the successful passage of sperm through the egg membrane. Kodama et al.'s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/57C12N15/11C12N15/10
Inventor 傅洪拓卜宪飞乔慧吴滟龚永生
Owner FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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