碩士 / 國立屏東科技大學 / 動物疫苗科技研究所 / 100 / Nocardia spp. are Gram-positive bacteria and widespread in soil. Fish nocardiosis caused by Nocardia seriolae is one of the important diseases in aquaculture industry worldwide. In this study, we use molecular cloning method to produce the recombinant glyceraldehyde - 3 - phosphate dehydrogenase (GAPDH), an outer membrane protein of N. seriolae, and analyze the efficacy of this vaccine candidate. We also choose an inactivated bacterin combined with several different adjuvants to compare the effects of immune protection. The largemouth bass was used as an animal model for selecting highly virulent strains of N. serolae and preparing the killed bacterins. First, the virulent strain was enriched, formalin-killed and homogenized by microfluidizer (MF) as antigen. The GAPDH coding gene of this strain was amplified by PCR and protein expressed as recombinant GAPDH in Escherichia coli BL21 (DE3) system. On the other hand, sera from largemouth bass challenged with N. seriolae were collected and Western blotting assay revealed that GAPDH shared the common antigenicity with N. seriolae and it reacted with antigen at 43.2 kDa. Fish were challenged with N. seriolae at 4 weeks after immunization 4 weeks. RPS values of MF Ag +GAPDH+ISA 763A, MF Ag +ISA 763A, MF Ag + Aluminum gel and GAPDH+ISA 763A group challenged by N. seriolae were at 6.3%, 18.8%, 31.3% and 62.5%, respectively. The challenged largemouth bass showed resistance against artificial infections with N. seriolae following immunization with GAPDH 4 weeks later. These results suggest that protective immune responses against N. seriolae are induced in largemouth bass.
Identifer | oai:union.ndltd.org:TW/100NPUS5289018 |
Date | January 2012 |
Creators | Hao-en Hung, 洪浩恩 |
Contributors | Shih-Chu Chen, 陳石柱 |
Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
Language | zh-TW |
Detected Language | English |
Type | 學位論文 ; thesis |
Format | 58 |
Page generated in 0.002 seconds