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Method of increasing cell permeability to trehalose by recombinantly producing a trehalose transporter

机译:通过重组生产海藻糖转运蛋白提高细胞对海藻糖渗透性的方法

摘要

There are provided trehalose transporter gene and a method of introducing trehalose into cells by using the gene. Candidates for the trehalose transporter genes were searched in P. vanderplanki EST, resulting in being obtained cDNA designated as Tret1. Tret1 encodes a 504 amino acid protein with 12 trans-membrane structures. Tret1 expression was induced by desiccation stress and predominant in the fat body. Functional expression of TRET1 in Xenopus oocytes showed that transport activity was specific for trehalose and independent of extracellular pH and electrochemical membrane potential. The direction of transport of TRET1 was reversible depending on the concentration gradient of trehalose. Apparent Km and Vmax of TRET1 for trehalose were extraordinarily high values. These results indicate that TRET1 is a facilitated, high-capacity trehalose-specific transporter. Tret1 is widespread in insects. Furthermore, TRET1 conferred trehalose permeability upon cells including those of vertebrates as well as insects.
机译:提供了海藻糖转运蛋白基因和使用该基因将海藻糖引入细胞的方法。在

中搜索海藻糖转运蛋白基因的候选者。 vanderplanki EST,得到的cDNA称为Tret1。 Tret1编码具有12个跨膜结构的504个氨基酸的蛋白质。 Tret1表达是由脱水压力诱导的,并且主要在脂肪体内表达。 TRET1在非洲爪蟾卵母细胞中的功能性表达表明,运输活性对海藻糖具有特异性,并且与细胞外pH和电化学膜电位无关。 TRET1的运输方向是可逆的,具体取决于海藻糖的浓度梯度。海藻糖的TRET1的表观Km和Vmax非常高。这些结果表明TRET1是一种便利的大容量海藻糖特异性转运蛋白。 Tret1在昆虫中很普遍。此外,TRET1使海藻糖对包括脊椎动物和昆虫的细胞具有渗透性。

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