Pérez-Arellano Isabel, Pérez-Martínez Gaspar
Departamento de Biotecnología, Instituto de Agroquímica y Tecnología de los Alimentos, Ap Correos 73, 46100 Burjassot, Valencia, Spain.
FEMS Microbiol Lett. 2003 May 16;222(1):123-7. doi: 10.1016/S0378-1097(03)00244-1.
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 degrees 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 microg of GFP per mg of total protein, from which 55% was fluorescent.
构建了一种用于干酪乳杆菌的表达载体，其含有可诱导的乳糖启动子和编码紫外可见绿色荧光蛋白（GFP(UV)）的基因作为报告基因。测定了干酪乳杆菌生长的不同条件，并对荧光以及合成的总蛋白进行了定量。维持中性pH对GFP(UV)表达的影响最大，其次是通气和30℃的温度。有利于GFP(UV)积累的环境因素与增强荧光的因素并不完全相关。因此，通过搅拌培养物进行充氧对荧光蛋白的比例影响最大，这与该蛋白的结构要求相符。获得的最高产量是每毫克总蛋白1.3微克GFP，其中55%是荧光性的。
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