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KMID : 0613820020120010096
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Journal of Life Science
2002 Volume.12 No. 1 p.96 ~ p.105
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Expression of an artificial gene encoding a repeated tripeptide lysyl-glutamyl-tryptophan in Tobacco plant
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Lee Soo-Young
Ra Kyung-Soo
Baik Hyung-Suk
Park Hee-Sung
Cho Hoon-Sik
Lee Yong-Se
Choi Jang-Won
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Abstract
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To investigate expression of the artificial gene encoding a repeated tripeptide lysyl-glutamyl-tryptophan in tobacco plant, the plant binary vector, pART404 has been constructed, which contains the duplicated CaMV 35S promoter, an artificial gene coding for repetitive polymer (Lys-Glu-Trp)64, and nopaline synthase (nos) terminator. The recombinant expression vector was introduced in Nicotiana tabacum (var. Xanthi) via Agrobacterium tumefaciens-mediated transformation. The transgenic calli selected by kanamycin containing medium were then regenerated to whole plants. Southern blot analysis indicated that five transgenic plants (No. 1, 7, 9, 43, 45) showed the hybridizing signals at 1.1 kb of the expected size on EcoRI digestion and each of the transgenic plants contained 1 or 3 copies of the artificial gene inserted into its genome. By northern blot analysis, the size of the hybridized total RNA was estimated to be approximately 1.2 kb and the RNA appeared generally to have the integrity. Western blot indicated that the protein was detected at the position of 33 kDa and the expression level of the polypeptide in the transgenic plant (No. 45) was measured to approximately 0.1% of the total protein.
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KEYWORD
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artificial gene, repetitive polymer (Lys-Glu-Trp)64, Agrobacterium tumefaciens, transgenic plant
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