Cloning and Expression of the Capsule Antigenic Gene from Erysipelothrix rhusiopathiae and Preparation of the Monoclonal Antibody

• Main Authors: Kue-Wan Zen, 鄭貴元
• Other Authors: Ming-Huei Liao
• Format: Others
• Language: zh-TW
• Published: 2008
• Online Access: http://ndltd.ncl.edu.tw/handle/81069332033699287274

碩士 === 國立屏東科技大學 === 生物科技研究所 === 96 === Erysipelothrix rhusiopathiae is a gram-positive bacterium that causes erysipelas in animal and erysipeloid in humans. Erysipelas in swine can occur as an acute septicemia or chronic disease,which is characterized by arthritis and endocarditis. The capsule of Erysipelothrix rhusiopathiae may be important in the pathogenicity of the disease. The objective of this study is to identify the antigenic regions of polypeptide of capsule for development of monoclonal antibody. The amino acid sequences with high antigenicity were predicated and amplified by PCR. The correct amplicons were cloned into pET-32a vectors, transformed into competent cell [ BL21 ( DE3 ) pLysS ], and expressed after induction with IPTG. BALB/c mouse were immunized with the expressed proteins purified with HiTrap affinity columns, sera from immunized mice were used to detect the proteins in Western blots. Then, the monoclonal antibody will be development and production.