Kolar M, Punt P J, van den Hondel C A, Schwab H
Institut für Biotechnologie, Mikrobiologie und Abfalltechnologie, Technische Universität Graz, Austria.
Gene. 1988;62(1):127-34. doi: 10.1016/0378-1119(88)90586-0.
An industrial Penicillium chrysogenum strain was transformed using two dominant selection markers, namely the bacterial gene for phleomycin resistance (ble) fused to a fungal promoter, and the acetamidase (amdS) gene from Aspergillus nidulans. Transformation frequencies of up to 20 transformants per microgram of DNA were obtained with the ble system. With the amdS marker the frequency was up to 120 transformants. Cotransformation was very efficient when using amdS as a selection marker. The introduction of pAN5-41B, a plasmid carrying the Escherichia coli lacZ gene fused to the strong glyceraldehyde-3-phosphate dehydrogenase gene (gpd) promoter from A. nidulans, resulted in the formation of blue colonies on XGal plates indicating expression of the lacZ fusion gene in P. chrysogenum. A more detailed analysis of expression levels in several transformants showed that up to 6% of the total amount of soluble protein consists of the beta-galactosidase fusion protein.
利用两个显性选择标记对一株产黄青霉工业菌株进行转化，这两个标记分别是与真菌启动子融合的抗腐草霉素细菌基因（ble）和来自构巢曲霉的乙酰胺酶（amdS）基因。使用ble系统时，每微克DNA的转化频率高达20个转化体。使用amdS标记时，频率高达120个转化体。当使用amdS作为选择标记时，共转化非常高效。导入携带与来自构巢曲霉的强甘油醛-3-磷酸脱氢酶基因（gpd）启动子融合的大肠杆菌lacZ基因的质粒pAN5-41B，导致在XGal平板上形成蓝色菌落，表明lacZ融合基因在产黄青霉中表达。对几个转化体中表达水平的更详细分析表明，可溶性蛋白总量的高达6%由β-半乳糖苷酶融合蛋白组成。
