A NOVEL TRANSFORMATION SYSTEM FOR PYRICULARIA-ORYZAE - ADHESION OF REGENERATING FUNGAL PROTOPLASTS TO COLLAGEN-COATED DISHES

被引：28

作者：

KIMURA, M ^{[1
]}

IZAWA, K ^{[1
]}

YONEYAMA, K ^{[1
]}

ARIE, T ^{[1
]}

KAMAKURA, T ^{[1
]}

YAMAGUCHI, I ^{[1
]}

机构：

[1] RIKEN, INST PHYS & CHEM RES, BIODESIGN RES GRP, WAKO, SAITAMA 35101, JAPAN

来源：

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY | 1995年 / 59卷 / 06期

关键词：

D O I：

10.1271/bbb.59.1177

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A new method for transformation of fungi to give efficient selection was developed. Protoplasts of Pyricularia oryzae, treated by PEG-CaCl2 with the blasticidin S (BS) deaminase gene (BSD) or the hygromycin phosphotransferase gene (hph), were transferred to liquid medium in collagen-coated dishes. They regenerated more efficiently in liquid than in agar, adhering to the collagen coating the dish. The colonies were continually kept under appropriate selective pressure by changing the medium. The BS resistant transformants of the isolate Hoku 1, well separated in the dish, were obtained at a frequency of 1000/mu g vector. In isolate P2, the frequency of transformation was 190/mu g vector with BSD and 4.5/mu g vector with hph. The value was 20 and 6 times, respectively, higher than that obtained by the conventional method using agar plates. The selected colonies on collagen were easily picked up by forceps, which allowed efficient replica-plating.

引用

页码：1177 / 1180

页数：4

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