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Cloning of Trehalose-6-Phosphate Phosphatase and Transformation to Tobacco

机译:海藻糖6-磷酸磷酸酶的克隆及其向烟草的转化

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摘要

The trehalose-6-phosphate phosphatase (TPP) is one of enzymes related to trehalose biosynthesis. But the researches of TPP were mainly focused in microorganism; few in the plant were reported. In this study, the TPP gene was cloned from Arabidopsis seedling by using RT-PCR (reverse transcription PCR) method. Then it was constructed to pET30a (+) vector and overexpressed in E.coli. Subsequently, the TPP was ligated into the plant expressional vector pBI121 and transformed to tobacco. Three transgenic individuals have obtained and one of them has transcribed successfully through Real-time QPCR detection. The trehalose content has less visible increase in P3 individual, but there was no obvious difference between the transgenic plant and control under salt stress.
机译:海藻糖6-磷酸磷酸酶(TPP)是与海藻糖生物合成有关的酶之一。但是TPP的研究主要集中在微生物上。据报道植物中很少。本研究采用逆转录PCR技术从拟南芥幼苗中克隆了TPP基因。然后将其构建为pET30a(+)载体,并在大肠杆菌中过表达。随后,将TPP连接到植物表达载体pBI121中并转化成烟草。已获得三名转基因个体,其中一名已通过实时QPCR检测成功转录。海藻糖含量在P3个体中几乎没有可见的增加,但是在盐胁迫下转基因植物和对照之间没有明显的差异。

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