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作 者 ：郭蓓;胡磊;何欣;陈雪梅*;蒋湘宁

期 刊 ：植物学通报 2008年 25卷 01期 页码：41-49

关键词：基因克隆与表达；定量PCR；胁迫耐受；转基因植物；TPS；

摘 要 ：海藻糖-6-磷酸合成酶(trehalose-6-phosphate synthase, TPS)是海藻糖合成途径中的一个关键酶, 近些年来受到广泛关注, 以往的研究多数集中于细菌和真菌等, 而对植物体内TPS基因的研究较少。为此, 我们首先通过RT-PCR(reverse transcription PCR)克隆得到了拟南芥的TPS基因, 进而将其构建到原核高效表达载体pET30a(+)上并在大肠杆菌BL21中进行高效诱导表达。在确认该基因能够在体外正常表达之后, 将其构建到植物表达载体上并转入烟草体内, 通过实时荧光定量PCR检测, 证明在转基因个体中目的基因已经在受体基因组上完成整合, 并且在其中的2个个体中实现了正常转录。在对目标个体进行海藻糖含量的测定以及抗逆性实验后发现, S5个体具有良好的表现, 其体内海藻糖的含量明显高于其它个体, 并具有较强的耐受盐胁迫的能力。

Abstract:Trehalose-6-phosphate synthase(TPS), which plays an important role in the synthesis of trehalose has received much attention in recent years. Research into TPS has mainly focused on bacteria and fungi. However, little research in plants has been reported. This paper describes the cloning and functional analysis of TPS from Arabidopsis. The Arabidopsis cDNA sequence of TPS in the pET30a(+) vector was heterogeneously overexpressed in BL21 of E. coli. Then, TPS was cloned into the plant expression vector pBI121 and transformed into tobacco. The expression of target gene in the transgenic tobacco plants was confirmed with real-time PCR. The transgenic tobacco line S5 showed higher trehalose level, which enhances the tolerance of transgenic plant to salt stress.

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