Iborra A, Sentandreu R, Gozalbo D
Departament de Microbiologia i Ecologia, Facultat de Farmàcia, Universitat de València, Burjassot, España.
Microbiologia. 1996 Sep;12(3):443-8.
Two plasmids (derived from YCplac22 and YEplac112) carrying a Candida albicans gene (including the 5' non-coding promoter sequences) coding for a 30 kDa membrane-bound protein, were used to transform Saccharomyces cerevisiae cells. A 30 kDa protein was immunodetected by Western blot in the membrane fraction of transformants. Northern analysis showed the presence of three mRNA species (of about 1.1, 0.7 and 0.5 kb) hybridizing with the C. albicans gene as a probe. The same result was obtained using the 5' and 3' regions of the gene as probes, whereas only a 1.1 kb mRNA was found in C. albicans and none was detected in S. cerevisiae control transformants. Thus, heterologous expression of this gene in S. cerevisiae results in a distinct pattern of mRNA processing, either due to the location on plasmid vectors and/or to differences in the mRNA processing systems in the two microorganisms.
携带编码一种30 kDa膜结合蛋白的白色念珠菌基因(包括5'非编码启动子序列)的两种质粒(源自YCplac22和YEplac112),用于转化酿酒酵母细胞。通过蛋白质免疫印迹在转化体的膜部分免疫检测到一种30 kDa的蛋白质。Northern分析显示存在三种与白色念珠菌基因杂交的mRNA种类(约1.1、0.7和0.5 kb)作为探针。使用该基因的5'和3'区域作为探针获得了相同的结果,而在白色念珠菌中仅发现了1.1 kb的mRNA,在酿酒酵母对照转化体中未检测到。因此,该基因在酿酒酵母中的异源表达导致了不同的mRNA加工模式,这可能是由于质粒载体上的位置和/或两种微生物中mRNA加工系统的差异所致。