| Summary: | 碩士 === 中興大學 === 分子生物學研究所 === 95 === Abstract
Rice is the major agricultural industry in Taiwan. The major course to developing and continuing the rice agriculture in Taiwan is to providing the additive values of domestic rice. Certainly, establishing the rice bioreactor system to producing high-value enzymes and proteins is a feasible way to advance the additive values of rice.
Transglutaminase (TGA) is an enzyme capable of stabilizing protein assemblies by gamma-glutamyl-epsilon-lysine crosslinks. The specific function of transglutaminase allows their biotechnological application in the foodstuffs industry: fish products (surimi), processed meat and sausages, chesses and yoghurt, ice creams, gelatines, chocolate etc. because food texture, firmness, elasticity, or fat and salt content can be modified. Several transglutaminase genes (tga) had been cloned from Streptomyces spp. by Dr. Ming-Te Young’s laboratory in Institute of Molecular Biology, National Chung Hsing University. The purpose of this study is to explore the possibility for overproducing the TGA in rice via transformed tga gene. We attempt to establish the bioreactor system by using the rice as a model plant to producing TGA.
In this study, the tga gene isolated from Streptomyces kentuckense was constructed into plant transformation vectors driven by CaMV 35S, rbcS, oleosin, and globulin promoter. The constructed genes were transferred into TN67 rice callus via Agrobacterium-mediated transformation. The regenerated plants were primary selected by G418. The results of PCR, Southern, RT-PCR and Western hybridization analysis indicated that the tga gene was present in the genome of transformed rice, and expressed tga mRNA and TGA protein with enzyme activity.
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