Quantitative aspects of the use of bacterial chloramphenicol acetyltransferase as a reporter system in the yeast Saccharomyces cerevisiae

被引:4
作者
Alipour, H [1 ]
Eriksson, P [1 ]
Norbeck, J [1 ]
Blomberg, A [1 ]
机构
[1] Univ Gothenburg, Dept Cell & Mol Biol, S-40530 Gothenburg, Sweden
关键词
CAT; yeast; immunological assay;
D O I
10.1006/abio.1999.4069
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The quantitative aspects of the use of the reporter system chloramphenicol acetyltransferase (CAT) has been evaluated in the yeast Saccharomyces cerevisiae. It was found that the CAT activity measured with a radiosotopic fluor diffusion assay was strongly dependent on the amount of yeast extract applied, both when CAT was expressed endogenously and when a purified Escherichia coli enzyme was investigated. Desalting the yeast extract by gel filtration partly eliminated the problem, indicating that some low-molecular-weight compound was involved in the phenomenon. However, the extract still exhibited stability problems on ice. An immunological CAT assay was tested and found to yield satisfactory quantitative result. (C) 1999 Academic Press.
引用
收藏
页码:153 / 158
页数:6
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