Platform formation for pharmaceutical glycoprotein production system in wood rot fungus

2017 Fiscal Year Final Research Report

• Project/Area Number: 15K14771
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Wood science
• Research Institution: Kyoto University
• Principal Investigator: Honda Yoichi 京都大学, 農学研究科, 教授 (70252517)
• Co-Investigator(Kenkyū-buntansha): 中沢 威人 京都大学, 農学研究科, 助教 (80608141)
• Research Collaborator: NGYUEN DongXuan 京都大学, 大学院農学研究科, 院生 ; SAKAGUCHI Taku 京都大学, 大学院農学研究科, 院生 ; NISHISAKA Emi 京都大学, 大学院農学研究科, 院生 ; KODERA Rina 京都大学, 大学院農学研究科, 院生 ; SAKA Chinami 京都大学, 農学部森林科学科, 学生
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: 白色腐朽菌 / 異種発現 / 糖タンパク質 / 遺伝子組換え

Outline of Final Research Achievements

With a transient gene expression system, that can evaluate gene expression level without nuisance of different gene dosage of integrated transgenes and their positional effect on the chromosome, it was demonstrated that a 14-bp stretch play a critical role in basic promoter function for beta-tubulin encoding gene in a white-rot fungus, Pleurotus ostreatus. The similar sequence was found among various basidiomycetes, suggesting that this stretch may serve as entry site for transcriptional factors as TATAA sequence does. A CT-rich sequence was also demonstrated to be important for the promoter function. Furthermore, a fusion gene encoding N-terminal of versatile peroxidase combined with luciferase was constructed and introduced in P. ostreatus. Activity of luciferase was detected in culture filtrate of this transformant, indicating that a reporter gene construct to investigate secretion system in wood-rot fungi was developed for the first time.

Free Research Field

森林生化学