Optimization of the green fluorescent protein (GFP) expression from a lactose-inducible promoter in Lactobacillus casei

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• 作者：Pé ; rez-Arellano ; Isabel ; Pí ; nez ; Gaspar
• 关键词：Green fluorescent protein ; Expression vector ; Lactose promoter ; Lactobacillus casei
• 刊名：FEMS Microbiology Letters
• 出版年：2003
• 期刊代码：34_03781097
• 类别：imm
• 出版时间：May 16, 2003
• 卷：222
• 期：1
• 页码：123-127
• 文件大小：250 K

摘要

An expression vector for Lactobacillus casei has been constructed containing the inducible lac promoter and the gene encoding ultraviolet visible green fluorescent protein (GFP_UV) as reporter. Different conditions to grow L. casei were assayed and fluorescence as well as total protein synthesized were quantified. The maintenance of neutral pH had the greatest incidence on GFP_UV expression, followed by aeration and a temperature of 30°C. Environmental factors favoring GFP_UV accumulation did not exactly correlate with those enhancing fluorescence. Therefore, oxygenation, by stirring the culture, had the greatest influence on the proportion of fluorescent protein, which is in accordance with the structural requirements of this protein. The highest yield obtained was 1.3 μg of GFP per mg of total protein, from which 55%was fluorescent.