Method for detecting drift treatment of target gene

A detection method and gene technology, applied in the field of gene drift processing and detection, can solve the problems that it is difficult to avoid the participation of exogenous gene drift, and achieve the effect of avoiding drift and ensuring environmental safety

Inactive Publication Date: 2011-06-08
NORTHWEST A & F UNIV
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Problems solved by technology

[0004] The purpose of the present invention is to provide a method for detecting the drift of the target gene, which solves th

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  • Method for detecting drift treatment of target gene
  • Method for detecting drift treatment of target gene
  • Method for detecting drift treatment of target gene

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[0042] 1 Scope

[0043] Including human β-defensin-3 gene cattle and sheep cell culture fluid, nuclear transfer operation fluid, reconstructed embryo culture fluid, and β-defensin-3 gene transgenic cattle and sheep offspring cell culture and nuclear transfer all liquids during operations Treatment and qualitative PCR detection of target gene drift.

[0044] It is suitable for the detection of gene drift in the production process of transgenic cows and goats derived from human defensin-3 genes.

[0045] 2 Terms, definitions and acronyms

[0046] 2.1 Target gene

[0047] The hBD3 gene is located in the p22-23 region of human chromosome 8 and includes 2 exons and 1 intron, with a total length of 1156bp.

[0048] 2.2 polymerase chain reaction (PCR)

[0049] The template gene sequence is first denatured by high temperature to become single-stranded. Under the action of DNA polymerase and suitable reaction conditions, two primers designed according to the template sequence are returned and com...

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Abstract

The invention discloses a method for detecting the drift treatment of a target gene. The method comprises the steps of: collecting liquids which possibly contain an exogenous gene such as transgenic cell culture solution, nuclear transplantation operation fluid, embryo culture solution and the like, performing high-pressure treatment, and then concentrating; and designing a primer for a gene sequence of the exogenous gene, specifically amplifying DNA fragments of the exogenous gene through a polymerase chain reaction (PCR) technology, and judging whether the exogenous gene still exists in the liquid produced during the screening of transgenic cells and the production and culturing of embryos and subjected to the treatment. The method for detecting the drift treatment of the target gene can completely avoid the drift of exogenous genes, and ensure the environmental safety during the production of transgenic animals. The method is applicable to the detection of genetic drift during the production of transgenic cows and goats derived from human defensin-3 genes.

Description

technical field [0001] The invention relates to a detection method for drift processing of target genes, in particular to a detection method for drift processing of target genes in the production process of hBD-3 gene cloned cattle or sheep. Background technique [0002] Transgenic cells and embryos contain exogenous target genes, and the drift of target genes in the process of cell culture and nuclear transfer is an important link in the safety evaluation of transgenics. It cannot have the drift of other exogenous genes, otherwise the drift of the target gene is not safe. The drift of the target gene refers to the process that the target gene undergoes polymerase chain reaction. Among them, the target gene-hBD3 gene is located in the p22-23 region of human chromosome 8, including 2 exons and 1 intron, with a total length of 1156bp; polymerase chain reaction (PCR) is It means that the template gene sequence is first denatured at high temperature to become a single strand, ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 华松张涌张辉王勇胜刘军权富生鲁成龙
Owner NORTHWEST A & F UNIV
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