Please use this identifier to cite or link to this item:
標題: 轉榖氨醯胺酵素(Transglutaminase, TGA)基因轉殖至大豆(Glycine max(L.) Merrill)之研究
Studies on the Transformation of the Transglutaminase Gene (tga) into Soybean (Glycine max(L.) Merrill)
作者: 施惠蓉
Hui-Jung Shih
Ming-Te Yang
Menq-Jiau Tseng
關鍵字: soybean;大豆;transglutaminase;bioreactor;轉榖氨醯胺酵素;生物反應器
Project: 興大園藝, Volume 31, Issue 3, Page(s) 57-72.
Horticulture NCHU, Volume 31, Issue 3, Page(s) 57-72.
大豆種子富含蛋白質,以大豆種子大量表現轉殖基因有極大潛力作為生物反應器,生產工業、食品、飼料及醫藥方面的產品的優勢。轉穀氨胺酵素(transglutaminase,TGA)會使蛋白濃縮液形成膠體化,因此在食品加工上有相當高的應用價值及潛力;例如:在漢堡、肉丸、魚漿、豆腐、植物蛋白粉末等可以改善其彈性、質地、口感、風味,並可增加儲存壽命。本研究乃嘗試將分離自Streptomyces ladakanum的tga基因(S.l-tga),分別黏接至水稻種子特有之啟動子鹽溶性球蛋白基因(globulin)及油膜蛋白基因(oleosin)之啟動子與CaMV35S及rbcS啟動子上,利用農桿菌基因轉移的方式,轉殖到'高雄選10號'大豆中。本研究之目的為建立以轉殖大豆種子為生物反應器之系統,並探討利用大豆生產轉穀氨胺酵素之可行性。
轉移四種構築載體之S.l-tga基因均可獲得再生植株。轉殖植株以PCR、南方墨點、KT-RCR及西方墨點雜交分析,其結果顯示tga基因已存在於轉殖之大豆植株基因粗中,且可正確表現tga RNA,並具有TGA酵素活性。TGA酵素活性分析之結果顯示轉殖大豆之葉片之TGA酵素活性最高為0.143 U/mg protein,為未轉殖對照組之4.9倍。

Soybean is a rich source of protein. Use transgenic soybean seedsas bioreactor to produce food, feed, industrial, and pharmaceutical proteins have the advantage over other bioreactor systems. Transglutaminase (TGA) is an enzyme capable of stabilizng protein assemblies by gamma-glutamy-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.
In this study, the tga gene isolated from Streptomyces ladakanum was constructed into plant transformation vectors driven by CaMV 35S, rbcS, oleosin, and globulin promoter. The constructed genes were transferred into soybean (Glycine max (L.) Merr. cv. KS10) cotyledonary node via Agrobacterium-mediated transformation. The regenerated plants were primary selected by kanamycin. The results of PCR, Southern, RT-PCR and Western hybridization analysis indicated that the tga gene was present in the genome of transformed soybean, and expressed tga mRNA and TGA protein with enzyme activity. The highest TGA activity in the leaves of tga-transformed soybean was 0.143 U/mg protein which was 4.9 folds of the controls.
ISSN: 0255-5921
Appears in Collections:第31卷 第03期

Files in This Item:
File Description SizeFormat Existing users please Login
興大園藝第三十一卷第三期p57-72.pdf4.62 MBAdobe PDFThis file is only available in the university internal network    Request a copy
Show full item record

Google ScholarTM


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.