Dieryck W, Lullien-Pellerin V, Marion D, Joudrier P, Gautier M F
Unité de Biochimie et Biologie Moléculaire des Céréales, INRA, Montpellier, France.
Protein Expr Purif. 1995 Oct;6(5):597-603. doi: 10.1006/prep.1995.1078.
The cDNA encoding a wheat (Triticum durum) lipid transfer protein of 9 kDa was inserted into an Escherichia coli expression vector, pIH902, and expressed in the bacteria as a fusion with the maltose binding protein. The fusion protein was then purified to homogeneity and subjected to factor Xa cleavage. Although complete cleavage of the fusion protein was obtained, the expected lipid transfer protein was not recovered; it appears to be degraded during protease digestion. However, a fluorescent lipid transfer assay demonstrated that the fusion protein has an activity identical to that of the wheat-purified lipid transfer protein. Thus, this expression system should allow further understanding of the structure/function relationships of wheat lipid transfer proteins.
将编码9 kDa小麦（硬粒小麦）脂质转移蛋白的cDNA插入大肠杆菌表达载体pIH902中，并作为与麦芽糖结合蛋白的融合蛋白在细菌中表达。然后将融合蛋白纯化至同质，并进行因子Xa切割。虽然获得了融合蛋白的完全切割，但未回收预期的脂质转移蛋白；它似乎在蛋白酶消化过程中被降解。然而，荧光脂质转移测定表明，融合蛋白具有与从小麦中纯化的脂质转移蛋白相同的活性。因此，该表达系统应有助于进一步了解小麦脂质转移蛋白的结构/功能关系。
