Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/88599
標題: 轉殖改造的大豆 β-伴球蛋白 β-亞基基因(β-Conglycinin β-Subunit)生產優質水產飼料用蛋白
Engineering of Soybean β-Conglycinin β-Subunit Genes for the Production of High Quality Soybean Proteins in Aquaculture Feeds
作者: Jun-Shian Yu
I-Hsiang Tsai
Ming-Te Yang
Wen-Hwei Hsu
Menq-Jiau Tseng
余俊賢
蔡奕祥
楊明德
許文輝
曾夢蛟
關鍵字: Soybean;β-Conglycinin β-Subunit;Aquaculture Feed;大豆;β-伴球蛋白β-亞基基因;水產飼料用蛋白
Project: 興大園藝, Volume 38, Issue 2, Page(s) 53-68.
摘要: 
Aquaculture is an important sector of fishery industry in Taiwan. Stable income of aquaculture is dependent on the price of fish meal. However, fish meal is in short supply and expensive, it is necessary to develop new product to substitute for the fish meal in aqua feeds. The high protein content, balanced amino acid composition, steady supply, and low cost of soybean made the derived products to be the ideal ingredients to replace the fish meal. The intrinsic limitations associated with soybean seed protein are low methionine, lysine content and poor palatability for fish. Attempts had been made to modify the soybean β-subunit of β-conglycinin gene (7Sb) by inserting the nucleotides of six and ten repeats of MGKMGR (7Sb-M56) and GMKGMR (7Sb-G510), respectively, so as to modify the amino acid composition of the soybean protein as a remedy. The modified 7Sb genes driven by the CaMV35S or Gm9 promoter (soybean seed-specific) had been sub-cloned onto pCambia 1304 vector. These plasmids were used for soybean transformation. Cotyledon nodes were used as explants for transformation of soybean [Glycine max (L.) Merr cv. 'Kaohsiung 10'] by Agrobacterium-mediated transformation. The transformed explants were selected with 100 ppm hygromycin. The regenerated plants and T1 progeny were examined by PCR, RT-PCR, and western blots. T1 progenies of 7Sb-G510 transgenic soybean were confirmed as evidenced by the presences of 7Sb-G510 gene, 7Sb-G510 mRNA, and 7Sb-G510 protein.

水產養殖是台灣相當重要的產業,魚粉的價格影響到養殖漁業的穩定收益。大豆的價格便宜及供應平穩,適合做為取代魚粉的原料。大豆種子富含有蛋白質及平衡的胺基酸組成分,其中 β-conglycinin 蛋白質含量占種子總蛋白的 35%。大豆蛋白與魚粉相較之下,缺乏胺基酸甲硫胺酸(methionine)及離胺酸(lysine),若添加甘胺酸(glycine)可以改善大豆適口性。本研究即是分別把含有六套 MGKMGR 及十套 GMKGMR 之重覆套數的核酸片段插入大豆 β-伴球蛋白(β-conglycinin)之 β-亞基 (β-subunit) (7Sb)基因,利用大豆作為生物反應器生產優質水產飼料用蛋白。本研究將二個改造之 β-亞基基因 (7Sb-M56 及 7Sb-G510),構築到含有CaMV35S 啟動子(持續性表現) 及 GmPM9 啟動子(種子專一表現) 的植物基因轉殖載體 pCambia 1304。轉殖載體以 mgfp 及 gusA 為報導基因,以 hptII 作為篩選基因,總共完成構築四種植物轉殖載體:pM56-1304-gus、pG510-1304-gus、pGm9-M56-1304-gus、 pGm9-G510-1304-gus。將四種植物轉殖載體利用農桿菌基因轉殖法轉殖到'高雄選 10 號'大豆子葉節。培殖體以 100 mg/L 的 hygromycin 進行篩選,並誘導再生。再生植株及 T1 後裔均經由 PCR、RT-PCR、西方墨點等方法檢測。目前已確認 T1 轉殖大豆品系攜帶有 7Sb-G510 基因,並表現 7Sb-G510mRNA 及 7Sb-G510 蛋白。
URI: http://hdl.handle.net/11455/88599
Appears in Collections:第38卷 第02期

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