Engineering of Soybean β-Conglycinin β-Subunit Genes for the Production of High Quality Soybean Proteins in Aquaculture Feeds

• Main Authors: Jun-Shian Yu, 余俊賢
• Other Authors: 曾夢蛟
• Format: Others
• Language: zh-TW
• Published: 2011
• Online Access: http://ndltd.ncl.edu.tw/handle/2kuea5

碩士 === 國立中興大學 === 分子生物學研究所 === 99 === Aquaculture contributes around 35% of the value of the Taiwan’s total seafood production, and is an important sector of fishery industry. Feed cost accounts 40% of the total production cost. 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, 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 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 which is anticipated to provide with stable supply of aquaculture meal thus benefiting farmers with better and stable income. Five plasmids had been constructed for soybean transformation, which were named pM56-1304-gus, pG510-1304-gus pGm9-M56-1304-gus, and pGm9- G510-1304-gus. 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 method. The transformed explants were selected with 100 ppm hygromycin. The regenerated plants and T1 progeny were examined by PCR, RT-PCR, and western blots. Twelve 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.