Site-directed knock-in plasmid vector for porcine mammary gland-specific high-expression lactoferrin gene and application thereof

Abstract

The invention relates to a site-directed knock-in plasmid vector for a porcine mammary gland-specific high-expression lactoferrin gene. With a 25251-25981 region of a reference sequence (GenBank:EU025875.1) of CSN1SI gene in the NCBI (National Center of Biotechnology Information) as a left homologous arm and a 25984-27112 region as a right homologous arm, a NheI restriction enzyme cutting site is introduced to the 5' end of the left homologous arm, a SalI restriction enzyme cutting site is introduced to the 5' end of the right homologous arm, a KpnI restriction enzyme cutting site is introduced to the 3' end of the right homologous arm, a T2A sequence and the CDS sequence of porcine LTF (lactoferrin) gene are inserted between the left homologous arm and the right homologous arm, and thereby a key sequence of a homologous recombinant site-directed inserted fragment is formed. The site-directed knock-in plasmid vector can be directly applied to construct genetically modified pigs with certain disease resistance against bacterial diarrhea, and particularly can enhance the resistance of piglets against the bacterial diarrhea.

Description

technical field [0001] The invention relates to the technical field of transgenic animal (pig) breeding, in particular to a site-specific knock-in plasmid vector for pig mammary gland-specific high-expression lactoferrin gene and application thereof. Background technique [0002] Bacterial diarrhea is an important cause of death in suckling piglets and weaned piglets. For a long time, the frequency of bacterial diarrhea in my country's pig industry is high and the scope is wide, resulting in low survival rate of piglets, low feed remuneration, low PSY of the whole group, slow growth and even stagnation, resulting in huge economic losses; at the same time, to a certain extent It has caused food and drug safety problems such as the abuse of veterinary antibiotics, excessive antibiotic residues, and drug resistance, as well as ecological and environmental pollution problems, posing a huge threat to ecological balance and human health. [0003] Lactoferrin (LTF) can optimize the...

AI Technical Summary

Problems solved by technology

For a long time, the frequency of bacterial diarrhea in my country's pig industry is high and the scope is wide, resulting in low survival rate of piglets, low feed remuneration, low PSY of the whole group, slow growth and even stagnation, resulting in huge economic losses; at the same time, to a certain extent It has caused food and drug safety problems such as the abuse of veterinary antibiotics, excessive antibiotic residues, and drug resistance, as well as ecological and environmental pollution problems, posing a huge threat to ecological balance and human health.

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