Gene transfer system and method for specific poultry primordial germ cell

A primordial germ cell and gene transfer technology, applied in biochemical equipment and methods, genetic engineering, microorganisms, etc., can solve problems such as low efficiency of gonadal gene transfer, transposon system without germ cell specificity, and many chimeras. , to achieve the effect of improving the efficiency of transgenic poultry, high gene transfer efficiency and strong specificity
CN105524941AInactive Publication Date: 2016-04-27YANGZHOU UNIV
2 Cites 5 Cited by

Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
YANGZHOU UNIV
Publication Date
2016-04-27
Estimated Expiration
Not applicable · inactive patent

Smart Images

  • Figure 1
    Figure 1
  • Figure 2
    Figure 2
  • Figure 3
    Figure 3
Patent Text Reader

Abstract

The invention provides a gene transfer system and method for a specific poultry primordial germ cell. The gene transfer system comprises a transposase helper plasmid containing a vasa promoter with a nucleotide sequence shown in SEQ ID NO.1. The gene transfer method comprises the steps of (1) constructing the transposase helper plasmid containing the vasa promoter or containing the vasa promoter and a universal enhancer element by using a bioengineering method; (2) constructing a transposon donor plasmid containing a target gene expression box required to be transferred by using the bioengineering method; (3) extracting the plasmids, uniformly mixing the transposon donor plasmid with the transposase plasmid according to a ratio of 1 to 2, and adjusting the final concentration of a mixed plasmid to be 500 ng-1.5 mu g / mu L; and (4) wrapping the mixed plasmid obtained in the step (3) with an embedding agent and transfecting PGCs cells in early embryo blood of poultry. Through the gene transfer system and method, efficient specific integration of exogenous genes in PGCs genomes in embryos of the poultry can be realized, so that the purpose of improving preparation efficiency of transgenetic poultry is achieved.
Need to check novelty before this filing date? Find Prior Art

Description

technical field

[0001] The invention belongs to the field of animal genetic engineering and relates to the establishment of a gene transfer system specific to poultry primordial germ cells and a poultry transgenic method mediated based on the gene transfer system. Background technique

[0002] Transposons are a type of DNA sequence that can freely jump (replicate or translocate) on the genome. It was first discovered in corn by Mc.Clintock in 1940, and later found in various organisms such as bacteria, fungi, and insects. . In recent years, transposon systems have been applied to non-virus-mediated transgenesis, especially in transgenic mammals. However, in the preparation of transgenic poultry, based on the special physiological and reproductive characteristics of poultry, that is: the early development of embryos has already started in the reproductive system of the female poultry, so even the newly laid eggs have developed into about 60,000 morphologically The blastodis...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More