萝卜抗真菌蛋白在酵母中的功能性表达。

Expression of functional Raphanus sativus antifungal protein in yeast.

作者信息

Alves A L, De Samblanx G W, Terras F R, Cammue B P, Broekaert W F

机构信息

F.A. Janssens Laboratory of Genetics, Catholic University of Leuven, Heverlee, Belgium.

出版信息

FEBS Lett. 1994 Jul 18;348(3):228-32. doi: 10.1016/0014-5793(94)00597-4.

DOI:10.1016/0014-5793(94)00597-4

PMID:8034047

Abstract

Rs-AFP2 is a 51 amino acid cysteine-rich peptide isolated from radish (Raphanus sativus) seeds that exhibits potent inhibitory activity against filamentous fungi. A cDNA clone encoding the Rs-AFP2 preprotein was modified by recombinant DNA methods to allow expression in the yeast Saccharomyces cerevisiae. This peptide was expressed in yeast as a fusion protein carrying at its N-terminus the prepro-sequences derived from the precursor of the yeast pheromone mating factor alpha 1. These sequences allow secretion of the biologically active peptide in a correctly processed form. Deletion of the mating factor alpha 1 pro-peptide drastically reduced the expression level of the peptide.

摘要

Rs-AFP2是一种从萝卜（Raphanus sativus）种子中分离出的富含51个氨基酸的半胱氨酸肽，对丝状真菌具有强大的抑制活性。通过重组DNA方法对编码Rs-AFP2前体蛋白的cDNA克隆进行修饰，使其能够在酿酒酵母（Saccharomyces cerevisiae）中表达。该肽在酵母中作为融合蛋白表达，其N端携带来自酵母信息素交配因子α1前体的前原序列。这些序列允许以正确加工的形式分泌生物活性肽。缺失交配因子α1前肽会大幅降低该肽的表达水平。