Method for preparing swine N-acetglutamic acid synthetase polyclonal antibody and application thereof

Abstract

The invention discloses a preparation method and application of a porcine N-acetylglutamate synthase polyclonal antibody. The steps are: cloning porcine NAGS gene, designing and synthesizing primers according to the gene sequence, and expressing the obtained full-length porcine The recombinant prokaryotic expression plasmid pLM1-NAGS of N-acetylglutamic acid synthase was transformed into Escherichia coli BL21; the BL21 of the transformed recombinant plasmid was cultured to an OD value of 0.4; IPTG 0.2mM was added to the cultured Escherichia coli BL21 to induce; The induced bacteria were collected by centrifugation, resuspended with PBS, lysed by adding lysozyme, and then sonicated. N-acetylglutamate synthase was used as an antigen to immunize Japanese large-eared white and black-eyed rabbits to obtain polyclonal antibody to N-acetylglutamate synthase, which was enhanced with incomplete Freund's adjuvant; the corresponding antiserum was obtained. The invention can detect the changes of distribution and expression activity of NAGS in suckling piglets at different developmental stages.

Description

technical field [0001] The invention relates to a preparation method and application of a porcine N-acetylglutamate synthetase polyclonal antibody. Background technique [0002] It has long been believed that pig milk can provide sufficient amino acids to meet the growth needs of suckling piglets, but research in recent years has shown that when pig milk is the only source of dietary protein for piglets, suckling piglets cannot grow optimally Performance, the research results show that the supply of arginine in breast milk and the endogenous synthesis of arginine in piglets at the age of 7-21 days is one of the main factors that limit the optimal growth of piglets, so increasing the supply of arginine in suckling piglets has Helps to maximize the growth potential of newborn piglets. [0003] N-acetylglutamate (NAG) is an allosteric activator of the first synthetase carbamoyl phosphate synthetase (CPSI) in the endogenous synthesis pathway of arginine (Adadjieva et al., 2001)...

AI Technical Summary

Problems solved by technology

In the small intestinal mucosa and mitochondria of the liver of suckling piglets, the lack of N-acetylglutamic acid synthetase leads to insufficient synthesis of N-acetylglutamic acid, thereby reducing the synthesis of intestinal citrulline and arginine, resulting in lactation Arginine deficiency in piglets

Although adding arginine to milk replacement diets for suckling piglets can greatly improve their performance, it is not suitable for actual pig production due to the high cost of fully automatic artificial feeding systems

Images