轉麩氨醯胺酵素基因轉移至水稻之研究

Studies on the Transformation of Transglutaminase Gene into Rice (Oryza sativa L.)

楊育臻(Yu-Chen Yang) ； 楊明德(Ming-Te Yang) ； 曾夢蛟(Menq-Jiau Tseng)

《植物種苗》 7卷4期 (2005/12) Pp. 1-20

https://doi.org/10.30077/SN.200512.0001

摘要

本研究已完成將Streptoverticillium mobaraense中篩選出的tga基因構築到植物轉殖載體，利用農桿菌法轉移將tga基因轉移到‘台農67號’水稻成熟胚所誘導之癒傷組織。在水稻所有組合之基因轉殖，均能獲得抗kanamycin及G418之再生水稻植株。轉殖植株以南方墨點、北方墨點及西方墨點雜交分析分析之結果顯示tga基因已存在於轉殖之水稻的基因組中，且可正確表現tga mRNA，並具有TGA酵素活性。TGA酵素活性分析之結果顯示轉殖水稻之葉片TGA活性最高為0.37U/mg protein，種子TGA活性最高為3.1 U/mg protein。以掃瞄式電子顯微鏡(SEM)觀察水稻種子，發現與未轉殖對照組沒有顯著差異，顯示tga基因轉移之水稻種子具有正常之外表及形態。以PCR分析T1代之tga基因轉殖水稻葉片，顯示tga基因可遺傳到下一代。

關鍵字

水稻 ； 基因轉移 ； 轉麩氨醯胺酵素

並列摘要

The purpose of this study was to explore the possibility for overproducing the TGA in rice via over-expressing the transglutaminase genes (tga). The tga genes isolated from Streptoverticillium mobaraense were constructed into plant transformation vectors driven by CaMV 35S, rbcS, oleosin, and globulin promoter. The constructed genes were transferred into callus of rice (Oryza sativa L. cv. Tinung 67) via Agrobacterium-mediated transformation. The regenerated plants were primary selected by kanamycin and G418. The results of PCR, Southern, Northern, and Western hybridization analysis indicated that the tga gene was present in the genome of transformed rice, and expressed tga RNA and TGA with enzyme activity. The highest TGA activity in the leaves and seeds of tga-transformed rice was 0.37 and 3.1 U/mg protein, respectively. There were no changes in the appearance and morphology in the seeds of transformed rice after observed with scanning electron microscope. The transformed tga gene could be inherited to the T1 progeny of transgenic rice.

並列關鍵字

rice ； gene transfer ； transglutamiase