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用于控制家禽球虫病的配子体抗原EmGam56的克隆、表达及纯化

Cloning, expression and purification of gametocyte antigen-EmGam56 for control of poultry coccidiosis.

作者信息

Ramalingam Vijayashanthi, Muthusamy Raman, Bohra Kasthuri, Palavesam Azhahianambi, Gopal Dhinakarraj

机构信息

Translational Research Platform for Veterinary Biologicals (TRPVB), Centre for Animal Health Studies, Tamil Nadu Veterinary and Animal Sciences University, Madhavaram Milk Colony, 600051 Chennai, India.

Department of Microbiology, Centre for Infectious Diseases, Saveetha Dental College and Hospitals, Saveetha Institute of Medical and Technical Sciences, Chennai, 600077 India.

出版信息

J Parasit Dis. 2023 Dec;47(4):773-777. doi: 10.1007/s12639-023-01610-w. Epub 2023 Aug 12.

Abstract

Poultry coccidiosis is an important devitalizing enteric protozoan disease caused by a group of obligatory intracellular apicomplexan parasites of the Genus contributing to major economic loss in commercial poultry worldwide. As the current method of chemotherapeutic control using ionophores in feed had led to development of drug resistant isolates, the need for development of prophylactic vaccines is the most viable alternate and eco-friendly control strategy as on date. Of the several candidate vaccines, the Gam 56 is one of the most promising candidates which protect the birds against , and , the three most pathogenic coccidian species infecting commercial chicken. Gam56 is a major wall forming component of macrogametocyte of and a candidate with high immunogenicity and low virulence. The present study was planned and carried out for the generation of expressed recombinant gametocyte antigen-Gam56 using pET 28(a+) as cloning vector and BL21 DE3 (pLysS) as prokaryotic expression system in a Bio-fermentor (New Brunswick™ Scientific BioFlo 310). The recombinant protein was purified by conventional (Ammonium sulphate precipitation) and by automatic purification system (AKTA prime) in Ni-NTA column for a planned immunization trial with experimental chickens.

摘要

家禽球虫病是一种由艾美耳属一群专性细胞内顶复门寄生虫引起的重要的使家禽衰弱的肠道原生动物疾病,在全球范围内给商业家禽养殖造成重大经济损失。由于目前在饲料中使用离子载体进行化学治疗控制的方法已导致耐药菌株的出现,因此开发预防性疫苗是目前最可行的替代方法和生态友好型控制策略。在几种候选疫苗中,Gam 56是最有前景的候选疫苗之一,可保护家禽抵御感染商业鸡的三种最具致病性的球虫种类——柔嫩艾美耳球虫、毒害艾美耳球虫和堆型艾美耳球虫。Gam56是柔嫩艾美耳球虫大配子体的一种主要的壁形成成分,是一种具有高免疫原性和低毒力的候选疫苗。本研究计划并实施,以pET 28(a+)作为克隆载体,BL21 DE3 (pLysS)作为原核表达系统,在生物发酵罐(新不伦瑞克™科学公司BioFlo 310)中表达重组配子体抗原-Gam56。重组蛋白通过常规方法(硫酸铵沉淀)和自动纯化系统(AKTA prime)在镍-亚氨基二乙酸柱上进行纯化,用于对实验鸡进行计划中的免疫试验。

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