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Idiosyncratic primer of repetitive sequence in gamma chromosome of oxen, and PRC method for identifying sex in early embryos of oxen

A technology of repetitive sequences and chromosomes, applied in the field of molecular biology, can solve the problems of pollution, false positives, complex identification methods, etc., and achieve the effect of low homology, strong specificity, and eliminating false negative errors of PCR

Active Publication Date: 2005-11-16
石家庄天泉良种奶牛有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the core sequence of the SRY gene has high homology (more than 80%) among mammals, such as 84% ​​homology with the core sequence of the SRY gene of people, so when carrying out sex identification of bovine embryos, the male operator The DNA of humans and other animals is likely to contaminate the identification process, especially when performing PCR operations in the field, due to environmental factors, it is easy to produce false positives, thus affecting the accuracy of embryo sex identification
[0005] my country's patent application (application number: 02111848.5) "Bovine Y-chromosome-specific primers and its application in bovine embryo sex identification", the invention uses nested primers, and its nested PCR adopts two-step reactions, and the identification method is relatively complicated. The embryos used still need to extract DNA, so it takes a long time and is very inconvenient

Method used

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  • Idiosyncratic primer of repetitive sequence in gamma chromosome of oxen, and PRC method for identifying sex in early embryos of oxen
  • Idiosyncratic primer of repetitive sequence in gamma chromosome of oxen, and PRC method for identifying sex in early embryos of oxen
  • Idiosyncratic primer of repetitive sequence in gamma chromosome of oxen, and PRC method for identifying sex in early embryos of oxen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Synthesis of specific primers

[0035] According to the Y-chromosome repetitive DNA sequence provided by Genbank, the present invention designs and synthesizes specific primers as follows:

[0036] Primer P1: 5'-TGATTGTTGATCCCCACAGAAGG-3'

[0037] Primer P2: 5'-TGAACTTTCAAGCAGCTGAGG-3'

[0038] A biotechnology company was entrusted to artificially synthesize two short oligonucleotide fragments, P1 and P2, on a gene synthesizer according to the primer sequences provided above for PCR reactions.

Embodiment 2

[0039] Embodiment 2: the synthesis of internal standard primer

[0040] The present invention designs and synthesizes a pair of primers according to the autosomal DNA sequence of cattle, and its sequence is as follows:

[0041] Primer sequence

[0042] 1 5'-TGAGGCATGGAACTCCGCTTG-3'

[0043] 2 5'-GGTGGTTCCACATTCCGTAGG-3'

[0044] A biotechnology company was entrusted to artificially synthesize two short fragments of oligonucleotides 1 and 2 on a gene synthesizer according to the primer sequences provided above for PCR reactions.

Embodiment 3

[0045] Embodiment 3: bovine embryo gender identification PCR method

[0046] 3.1 Carry out gender identification through bovine peripheral blood genomic DNA, and detect the specificity of primers.

[0047] First collect 5mL of peripheral blood from male and female dairy cows respectively, take 1mL of whole blood and use a kit to extract genomic DNA, dissolve it in 200 μL of sterilized ultrapure water, and take 2ng for multiplex PCR amplification.

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Abstract

A primer specific to the repetitive sequence of ox Y-chromosome has a sequence 5'-TGATTGTTGATCCCACAGAAGG-3',5'-TGAACTTTCAAGCAGOTGAGG-3'. In multiple PCR test, said specific primer and internal standard primer are used to amplify 131 bp specific fragment of Y-chromosome and 250 bp fregment of internal standard gene for bull or only 250 bp fragment of internal standard gene for cow, so it can be used to determine the sex of early ox embryo.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a specific primer sequence of bovine Y-chromosomal repeat sequence for bovine early embryo sex identification by PCR method. Background technique [0002] In the field of agricultural biotechnology, controlling the sex of livestock has been a dreamed aspiration for many years. Because through sex control, some sex-restricted traits (such as cow lactation) and sex-affected production traits (meat production, hair production) can obtain greater economic benefits in more ideal sexes. Increase the selection intensity in breeding to obtain the maximum genetic progress, and at the same time exclude harmful genes in livestock and poultry populations. From the perspective of research and development, the methods of gender control are basically divided into two aspects: one is the separation technology of X and Y sperm, combined with artificial insemination, to control the sex of offspri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 张莉余文莉
Owner 石家庄天泉良种奶牛有限公司
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