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首页> 外文期刊>Asia-Pacific Journal of Science and Technology >Production and detection of - glucuronidase (GUS) protein in fruit tissue of papaya (Carica papaya L.) and banana (Musa acuminata L.) using Agrobacterium transient transformation
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Production and detection of - glucuronidase (GUS) protein in fruit tissue of papaya (Carica papaya L.) and banana (Musa acuminata L.) using Agrobacterium transient transformation

机译:农杆菌瞬时转化法在番木瓜(Carica papaya L.)和香蕉(Musa acuminata L.)的果实组织中生产和检测-葡萄糖醛酸酶(GUS)蛋白

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摘要

Heterologous protein production and determination of fruit specific promoter require gene transformation into target plant host. Fruits transformed gene by Agrobacterium is the convenient and rapid method. The outcome is clearly revealed by producing heterologous and functional protein. In this study, two fruit plant species for heterologous protein production, banana and papaya were transiently transformed with In this study, two fruit plants; banana ((Musa acuminata L.) and papaya (Carica papaya L.) were determined the GUS gene expression and heterologous protein production after transiettly transformation with Agrobacterium tumefaciens AGL-1 containing a plant expression vector, pCAMBIA1304 with?-glucuronidase (GUS) gene as a reporter gene during mature fruit and color break fruit . Factors influencing transformation efficiency including fruit developmental stages, bacterial concentration and incubation time were investigated. The transformation efficiency was determined by GUS staining and GUS enzymatic activity. The results showed that bacterial dilution at 1:10 for 30 min, was exhibited the most effective results in both fruit. This method was also applicable to tomato, a fruit model commonly used for gene expression, promoter and protein production.
机译:异源蛋白质的生产和果实特异性启动子的确定需要将基因转化为目标植物宿主。农杆菌转化的果实基因是一种方便快捷的方法。通过产生异源和功能蛋白清楚地揭示了结果。在本研究中,使用异种植物瞬时转化了两种用于生产异源蛋白质的果树物种香蕉和木瓜。用含有植物表达载体的农杆菌AGL-1,带有β-葡萄糖醛酸苷酶(GUS)基因的pCAMBIA1304瞬时转化后,确定了香蕉((Musa acuminata L.)和番木瓜(Carica papaya L.)的GUS基因表达和异源蛋白质产生。作为成熟果实和断色果实的报道基因,研究了影响果实转化阶段,细菌浓度和培养时间的因素,并通过GUS染色和GUS酶活性确定了转化效率,结果表明细菌稀释度为1。在10:30的30分钟中,这两种水果均显示出最有效的结果,该方法也适用于番茄,番茄是一种通常用于基因表达,启动子和蛋白质生产的水果模型。

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