为了研究GO基因在植物中的功能,获得转基因植株,用限制性内切酶将乙醇酸氧化酶(GO)基因从克隆载体pBSSP上切下,连接到植物双元表达载体pBin117的CaMV 3S启动子和NOS 终止子之间,成功构建了GO基因植物表达载体pBinGO.在此基础上以嘎拉苹果叶片为受体,通过根癌农杆菌介导法将GO基因导入苹果,获得了2个抗性芽,PCR检测初步表明GO基因已整合到嘎拉苹果基因组中.
In order to study the gene function of GO,obtain the transgenic plant,a plant expression vector pBinG O was constructed by recombinating the G lylate oxidase gene fragment from cloning vector pBSSP5 and the binary vector pBin117 at the EcoR Ⅰsite.The fragment of G lylate oxidase gene was inserted between of CaMV 35Spromotor and NOS terminator in pBin117.The identification results showed that the plant expression binary vector pBinG O was constructed successfully.Subequentely GO gene was introduced into apple leaves mediated by Agrobacterium tumerfaciens.Two transgenic resistant plants were proved by PCR preliminarily that the GO gene had integrated into Apples genome.
[1] Moran J F,Becana M.Drought induces oxidative stressing pea plants[J].Planta,1994,194:346-352.
[2] Esquivel M G,Ferreira R B,Teixeira A R.Protein degradation in C3 and C4 plants with particular reference to ribulose bisphosphate carboxylase and glycolate oxidase[J].J Exp Bot,1998,49:807-816.
[3] Zelitch I.Increased rate of net photosyhthetic carbon dioxide uptake caused by the inhibition of glycolate oxidase[J].Plant Physiol,1966,41:1623-1631.
[4] 孟庆伟,赵世杰,许长成,等.田间小麦叶片光合作用的光抑制和光呼吸的防御作用[J].作物学报,1996,22(4):470-475.
[5] Akiko K,Go T.Photorespiration protects C3 plants from photooxidation[J].Nature,1996,384:557-560.
[6] Dvir Taler,David Kenigsbuch.Plant eR genes that encode photorespiratory enzymes confer resistance against disease[J].The Plant Cell,2004,16:172-184.
[7] 刘进元,常智杰,赵广荣,等.分子生物学实验指导[M].北京:清华大学出版社,2002:132-133.
[8] 邓振山,赵立恒,王红梅.苹果常见主要真菌性病害的研究进展[J].安徽农业科学,2006,34(5):932-933,935.
[9] 潘增光,邓秀新.苹果组织培养再生技术研究进展[J].果树科学,1998,15(3):261-266.
[10] 王关林,方宏筠.植物基因工程原理与技术[M].北京:科学出版社,1998.
