| Summary: | 碩士 === 國立宜蘭大學 === 園藝學系碩士班 === 97 === Classical swine fever virus (CSFV) is the causative agent of classical swine fever (CSF), a highly contagious infection of swine. At present, vaccine is an effective method to prevent CSFV. However, traditional attenuated vaccines could not discriminate between vaccinated and infected animals. The E2 protein of CSFV contains major antigenic determinants that is conserved and involved in neutralization by antibodies. Heat stress in living cell causes multiple changes that ultimately affect membrane structure and function. It has been suggested that HSP function as molecular chaperons, which direct the proper folding of the protein, as well proper assembly of the protein complex.
We had constructed the surface structural protein gene of classical swine fever virus (E2 gene) or heat shock protein gene of low molecular weight (HLJ1 gene) with the promoter of CaMV 35S as plant transfer vectors. The constructed plasmids were transferred into soybean (‘Kaohsiung SEL.10’) and cabbage (‘K-Y cross’ and ‘Summer Summit’) via Agrobacterium mediated transformation. The regeneration rates of ‘K-Y cross’ cabbage, ‘Summer Summit’cabbage and soybean were 0.08~0.13%, 0.1~0.2% and 1~2.5%, respectively. The transformed plants were examined by PCR and RT-PCR. Six and two independent transgenic ‘K-Y cross’ lines were regenerated, integration of the E2 and HLJ1 gene were confirmed by PCR (66.7% and 50% positive reaction) and specific transcripts of the expected molecular size were detected by RT–PCR (75% positive reaction). One and three independent transgenic ‘Summer Summit’ lines were regenerated, integration of the E2 and HLJ1 gene were confirmed by PCR (100% and 33.3% positive reaction) and specific transcripts of the expected molecular size were detected by RT–PCR (100% positive reaction). Twenty-one and five independent transgenic soybean lines were regenerated, integration of the E2 and HLJ1 gene were confirmed by PCR (52.4% and 80% positive reaction) and specific transcripts of the expected molecular size were detected by RT–PCR (100% positive reaction).
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