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KMID : 0578319910010030261
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Molecules and Cells
1991 Volume.1 No. 3 p.261 ~ p.266
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Synthesis of an Artificial Gene for the shorthron sculpin Antifred Protein and Its Introduction into Tobacco Plants
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Park, Yu-Shin
Chang, Seok-So/Park, Duch-Hoon/Kim, Dal-Ung/Nam, Hong-Gil
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Abstract
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Antifreeze protein (AFP) is a family of small proteins found in the polar fishes and some insects. AFPs appear to prevent ice formation in the blood at subzero temperatures, thus reducing freezing injury. To exploit the possibility of introducing freezing tolerance into plants, we synthesized an artificial gene which codes for a 45 amino acid antifreezer protein and introduced it into tobacco cells. The synthetic gene was designed to code for a Shorthorn sculpin AFP by assembling from 4 oligonucleotides. The synthetic gene inserted into a plant expression vector pGA643 was used to transform tobacco cells by the Agrobacteriummediated leaf disk transformation method. Transgenic plants were generated by kanamycin selection and introduction of the synthetic gene into the plant genome was confirmed by NPT II assay and Southern hybridization. Furthermore transcripts of the synthetic gene were detected by Northern blot analysis and production of a new small polypeptide was observed in the extract of the transgenic plants as monitored by polyacrylamide gel electrophoresis.
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