Cow fetus fibroblast cell containing Iprl macrophage metamerism expression vector

Abstract

The invention discloses a cow fetus fibroblast cell containing an Iprl macrophage metamerism expression vector. A cow fetus fibroblast cell line which contains an Iprl macrophage metamerism expression vector Psp-EGFP-Iprl, is based on the vector and contains the Iprl macrophage metamerism expression vector is established, and the cell line is used as a nucleus donor cell, which provides massy foundation for transgene cow with tuberculosis. In the invention, an exogenous expression vector Psp-EGFP-Iprl is transduced to the cow fetus fibroblast cell through an electrotransfection method, the expression condition of a marked gene EGFP is observed by a fluorescence microscope, positive cells are selected by G418, the target gene Iprl is confirmed to be integrated into the genome of the cow fetus fibroblast cell by PCR identification, and finally the cow fetus fibroblast cell subjected to the transfection of the Iprl gene is moved into cow denucleated oocyte as a donor to obtain a transgene clone embryo.

Description

technical field [0001] The invention belongs to the technical field of transgenic cloning animals, and relates to the construction of nuclear donor cells of transgenic cloned embryos, in particular to bovine fetal fibroblasts containing Ipr1 macrophage-specific expression vectors. Background technique [0002] Mycobacterium tuberculosis is the pathogenic bacterium of tuberculosis that is zoonotic. Mycobacterium tuberculosis is divided into three types: human type, bovine type and avian type, and there is a cross-infection phenomenon among the three. Mycobacterium bovis mainly infects cattle. Bovine tuberculosis is a zoonotic chronic infectious disease caused by Mycobacterium bovis. The diseased cattle are characterized by gradual emaciation and the formation of nodular granulation in tissues and organs. swelling and caseous necrosis. The cause of bovine tuberculosis is its easy infection and serious harm. Therefore, the World Organization for Animal Health lists it as a B-t...

AI Technical Summary

Problems solved by technology

However, long-term use of antibiotics will not only lead to drug resistance of bacteria, but also the problem of drug residues in milk, so it is urgent to seek a new method for the treatment of tuberculosis

[0004] With the development and improvement of molecular biology, molecular genetics and genetic engineering technology, new ideas have been provided for animal disease resistance breeding. In the 1980s, pronuclear injection was used to produce transgenic animals, but this method has random integration of foreign genes. Uncertainty in transmission of exogenous genes with gamete lines limits the further development of this technology

Somatic embryonic stem cells can replace pronuclear injection, but embryonic stem cells have not yet been established in livestock, so they cannot be widely used

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