Cloning and characterization of gag gene from Caprine arthritis-encephalitis virus (CAEV) as a tool for vaccine development

• Main Authors: Shin-Min Chang, 張詩敏
• Other Authors: Wen-Hsing Chang
• Format: Others
• Language: zh-TW
• Published: 2007
• Online Access: http://ndltd.ncl.edu.tw/handle/49979838834441195484

碩士 === 國立嘉義大學 === 農業生物技術研究所 === 95 === Caprine arthritis-encephalitis virus (CAEV) is a retrovirus belonging to the Lentivirus subfamily. CAE is a chronic goat disease characterized by progressive inflammation in multiple organs or tissue systems. CAEV’s transmission pathways can be either from dam to kid through colostrums or through prolonged contact between infected and healthy animals. The prevalence of CAEV infection, especially in zones of production, has significantly reduced farmers’ profits due to fall in production of goat milk, early slaughter of undesirable stock, and delay of exports. Currently, there is no commercially available vaccine to control or prevent CAEV infection. The CAEV’s gag gene has been shown to possess good antigenic epitopes and high conservative amino acid sequence in relative CAEV strains. Therefore, the gag gene product may serve well as subunit vaccine against CAEV infection in goat. In this report, CAEV’s gag gene was first amplified from infected goat milk by nested RT-PCR. The product of nested RT-PCR was confirmed by DNA sequencing analysis and found to encode a 996 bp open reading frame of CAEV’s gag gene. This cloned gag gene was applied in a M15 E. coli expression system via the vector of pQE-trisystem. By western blotting assay, the gag gene product is demonstrated to express in M15 after IPTG’s induction. The gene product of cloned gag gene was further purified by nickel-affinity chromatography to homogeneity. These results lay the foundation of developing subunit vaccine against CAEV in the near future.