以甘油醛-3-磷酸脫氫酶基因啟動子研究美白菇轉形系統

Abstract

隨著遺傳工程的發展，目前已有許多成熟的異源表達系統可表現外來基因，生產重組蛋白質。由於分子生物學與基因轉形技術愈趨成熟，菇類異源表達系統的研究也在近十年逐漸引起科學家的重視。 本研究以近年頗受大眾喜愛之美白菇作為研究材料，選殖此菇種的甘油醛-3-磷酸脫氫酶基因序列，再設計可擴增啟動子與其轉錄區域當中第一段內隱子之引子並釣取啟動子片段。以獲得之啟動子片段分別建構帶有潮黴素抗藥性基因-潮黴素磷酸轉移酶基因 (hygromycin phosphotransferase gene, hph) 之篩選標記載體，與帶有綠色螢光蛋白質基因 (enhanced green fluorescent protein, egfp) 或

As the progress of genetic engineering was developed, there were many mature heterologous expression system which could express heterologous genes and produce recombinant protein. Because of the maturing molecular biology and gene transformation technology, research on mushroom heterologous expression systems attracted scientist’ attention for the past decade. The goal of this study is to establish a transformation system of Hypsizygus marmoreus (variant), which is popular in recent years. After cloning the glyceraldehyde-3-phosphate dehydrogenase gene (gpd) sequence of H. marmoreus (variant), the primers were designed to amplify the sequence of the promoter with the first intron in the transcription region. We constructed two kinds of plasmids, one with the selectable marker, Escherichia coli hygromycin B phosphotransferase gene (hph), and the other with the reporter genes, ehanced green fluorescent gene (egfp) and