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Method for guiding exogenous substances into chick embryos

A technology of exogenous substances and chicken embryos, applied in the field of introducing new exogenous substances into chicken embryos, can solve the problems that the method is not uniformly and clearly described, it is difficult to repeat, and the process is complicated, so as to achieve good promotion and application value, easy operation, and operation easy effect

Pending Publication Date: 2018-02-23
YANGZHOU UNIV
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Problems solved by technology

The intravascular injection method has obvious advantages due to its high transfer efficiency of exogenous substances, but the method of intravascular injection has not been uniformly and clearly described, and multiple intravascular injection methods have disadvantages such as cumbersome operation, complicated process, and difficulty in repeating, which limit the quality of chicken embryos. Further application of vascular injection requires further optimization and improvement

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  • Method for guiding exogenous substances into chick embryos
  • Method for guiding exogenous substances into chick embryos
  • Method for guiding exogenous substances into chick embryos

Examples

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Embodiment Construction

[0021] A new method for introducing foreign substances into chicken embryos:

[0022] (1) Use tweezers to open the blunt end of the early embryo eggs (13-17HH) incubated for 48-58 hours under sterile conditions. The opening size is a round hole with a diameter of 1-1.5 cm. Find the embryo position under a stereo microscope. Use ophthalmic forceps to gently pry open the outer shell membrane to expose the embryonic blood vessels.

[0023] (2) The exogenous substances that need to be introduced (such as wrapped plasmid vectors, cells, etc.), in this example, the liposome-wrapped plasmid (pEGFP-N1) is used for injection, and the mass-to-volume ratio of liposomes and plasmids is 1:3, put the wrapped plasmid in a 37-degree incubator and incubate for 30mins, then use a micropipette to fill it into the glass injection needle prepared in advance, and inject it into the blood vessel of the chicken embryo through a microinjector. The parts are shown in the figure ( Figure 1a , Figure...

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Abstract

The invention discloses a method for guiding exogenous substances into chick embryos, and belongs to the technical field of the biology. The method specifically comprises the following steps: (1) forming an opening in an 13-17 HH early embryo egg which has been incubated for 48-58 h under the aseptic condition, finding out the position of an embryo, and stripping an embryo egg shell film to exposeembryo blood vessels; (2) injecting the exogenous substances required to be guided in into a glass entry needle, and then injecting the exogenous substances into the embryo blood vessels through a micro-injector; and (3) after injection is finished, adding mycillin into an injected part, and then sealing by using a medical adhesive tape. The exogenous substances are guided in through blood vesselinjection in a chick embryo early stage (13-17 HH), and thus, the method lays a foundation for extensive application of chick embryo injection. The method is suitable for guiding in chick embryos exogenous genes or cells, is suitable for preparation of transgenic chickens and chimera, and is simple, convenient and rapid to operate; and the exogenous substance guiding-in efficiency is high.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a new method for introducing exogenous substances into chicken embryos. Background technique [0002] As a classic model of embryonic biology, the chicken embryo has been used to explore embryonic development and embryonic cell differentiation, especially in medical models with broad application prospects. The introduction of exogenous substances into chicken embryos is of great significance to production research such as gene function verification, transgenic breeding and chimera preparation. Therefore, an efficient and simple method for introducing exogenous substances can not only provide technical support for chicken embryo development biology and transgenic chicken production, but also provide an experimental basis for related research. At present, there are a large number of reports on the method of chicken embryo injection both at home and abroad. At present, the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/89
CPCC12N15/89
Inventor 李碧春靳锴左其生张亚妮
Owner YANGZHOU UNIV
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