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Construction of Genetically Modified Tobacco (Nicotiana tabacum cv. Petit Havana) Harboring ompH(A:3) from Pasteurella multocida (A:3)
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  • Construction of Genetically Modified Tobacco (Nicotiana tabacum cv. Petit Havana) Harboring ompH(A:3) from Pasteurella multocida (A:3)
저자명
Jeong. Ki-Yong,Maeng. Kyung-Ah,Song. Ji-Hyun,Hong. Sung-Youl,Chung. Tae-Young,Kwon. Moo-Sik
간행물명
Biotechnology and bioprocess engineering
권/호정보
2008년|13권 1호|pp.115-121 (7 pages)
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한국생물공학회
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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Fowl cholera, caused by Pasteurella multocida (A:3), is a fearsome disease leading to a nonproductive influence upon poultry industry. It has been known that outer membrane protein H (OmpH) in the bacterium is a strong candidate to bring on the notorious ailment. Genetically modified (GM) tobacco (Nicotiana tabacum cv. Petit Havana) harboring ompH(A:3) was constructed to develop a plant expression system for the protein, OmpH(A:3). Some 987 bp-long (ORF with the stop codon, TAA) of the ompH(A:3) excluding the nucleotide for signal peptide, was amplified by RT-PCR with the gene specific primers and pGEM-T-ompH(A:3) as template DNA. The PCR-amplified DNA was ligated into BamHI/Sacl-cut pBI121 to obtain a recombinant plasmid, pBI121-ompH(A:3). It was then transformed into Agrobacterium tumefaciens (LBA 4404) by liquid nitrogen method to generate a recombinant clone of Agrobacterium LBA4404/pBI121-ompH(A:3). The Agrobacterium LBA4404/pBI121-ompH(A:3) was inoculated into leaf discs of tobacco (2 day old). The gene-transfected leaves were cultured on Murashige-Skoog basal medium containing kanamycin (50 mg/mL) to generate numerous calli, from which some GM tobacco plants were obtained. Transgenicity of the tobacco plant was confirmed by PCR screening along with the DNA sequencing. Also, its expression in the GM-tobacco was examined qualitatively as well as quantitatively by ELISA/Western blot. These results suggest that the genetically modified tobacco plant can be potentially used as a model system to develop plantbased vaccine against the fowl cholera.