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Spermary-specific protein gene sequence primer and non-electrophoresis method for detecting and identifying cow embryo gender

A specific protein and gene sequence technology, applied in the direction of DNA/RNA fragments, recombinant DNA technology, etc., can solve problems such as weakness, shorten the time, improve the sensitivity, and facilitate the promotion and application.

Inactive Publication Date: 2009-06-10
INST OF ANIMAL SCI & VETERINARY MEDICINE SHANDONG ACADEMY OF AGRI SCI
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Problems solved by technology

my country has established the PCR technology for bovine embryo sex identification by using the SRY gene and ZFY gene of cattle, but the work on the industrialization of this technology is still relatively weak

Method used

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  • Spermary-specific protein gene sequence primer and non-electrophoresis method for detecting and identifying cow embryo gender

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Put 3-4 drops (each drop 200 μl) of phosphate buffered saline (PBS) in a petri dish, and use MN-151 metal blades to cut the embryos under a stereomicroscope (SZX7-3121, OLYMPUS). About 10% of the inner cell mass of the morula or about 10% of the vegetative ectoderm of the blastocyst are cut for biopsy, and the cut embryos are to be transplanted. Clean the blade with 70% alcohol and sterilized ultrapure water every time an embryo is cut. Six embryos were divided and sampled according to the above method.

[0016] Wash the 6 divided embryo samples with phosphate buffered saline (PBS) for 3 times, respectively draw the divided embryo samples (total volume about 2 μl) with a pipette gun (specification 0.5-10 μl), and transfer them into 0.2ml PCR reaction tubes (6), each PCR reaction tube was pre-put 5 μl alkaline lysate (ALB) (ALB composition: 50mM DTT and 200mM KOH, that is, 50mM dithiothreitol and 200mM potassium hydroxide), 65 ℃ , 10min. Then 5 μl of neutralizing solu...

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Abstract

The invention discloses an atopic protein gene sequence primer of testis with sequence as 5'-CCGCCATTACGCCCCCGACTTG-3' and 5'-GGGCCGCTGTTCCTGCTCCTCAT-3', which is characterized by the following: adopting normal PCR method and non-gel electrophoresis to test early embryo sex of cow; improving sensitivity of reaction; shortening the identifying time without polluting; fitting for transplanting embryo.

Description

technical field [0001] The invention relates to a testis-specific protein gene sequence primer and a method for non-electrophoretic detection and identification of cow embryo sex. Background technique [0002] For a long time, people have longed to control the sex of domestic animals, especially for economically valuable domestic animals such as cattle, sheep, and pigs. Sex identification of early embryos, especially pre-implantation embryos, before transplantation has important scientific research value and commercial application value in animal husbandry production. The main methods for identifying the sex of embryos include: cytogenetic analysis, determination of X chromosome-associated enzyme activity, analysis of embryonic developmental rate differences, detection of male-specific antigens, and application of Y chromosome-specific DNA probes. Although X and Y spermatozoa have been successfully isolated using flow cytometry and commercially available semen is available,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 黄金明刘玉庆游伟张俊功谭秀文吴乃科朱荣生柳尧波武英张秀美
Owner INST OF ANIMAL SCI & VETERINARY MEDICINE SHANDONG ACADEMY OF AGRI SCI
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