I M Gerasymenko, M G Mazur, Y V Sheludko, N V Kuchuk
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引用次数: 0
摘要
将转基因植物作为生产具有生理活性的人类蛋白质的工厂引起了人们的特别关注,因为这些转基因偶尔逃逸到环境中可能会造成健康问题。在培育生产有药用价值蛋白质的植物品种的同时,还应该开发适合控制转基因行为的检测方法。我们在此介绍一种多重 PCR 方案,用于揭示已成功导入植物基因组的两个人类基因(编码生长激素和干扰素 alpha2b)。为检测人类生长激素编码序列而设计的引物对可以扩增人类基因组中全长基因和通常用于植物转化的无内含子编码序列的不同大小的片段。建议使用该引物对,以排除因样本受到人类 DNA 污染而导致的假阳性结果。在培育携带人类基因的转基因植物时,这种对照也可用于 PCR 分析。
MULTIPLEX PCR ASSAY FOR DETECTION OF HUMAN SOMATOTROPIN AND INTERFERON ALPHA2b GENES IN PLANT MATERIAL.
Using transgenic plants as factories for production of physiologically active human proteins arouses special concern because occasional escape of such transgenes into environment may cause health problems. Creation of plant varieties producing pharmaceutically valuable proteins should be accompanied by development of detection methods suitable for controlling the transgene behavior. Here we describe a multiplex PCR protocol for revealing of two human genes (encoding growth hormone and interferon alpha2b) that have been successfully introduced into plant genomes. The primer pair designed for detection of human growth hormone coding sequence amplifies fragments of different size from the full-length gene in the human genome and the intronless coding sequence usually used for plant transformation. Application of this primer pair may be recommended for ruling out false positive results due to sample contamination with human DNA. Such a control may be useful also in PCR analysis during establishing of transgenic plants carrying genes of human origin.