Graessle S, Haas H, Friedlin E, Kürnsteiner H, Stöffler G, Redl B
Institut für Mikrobiologie (Medizinische Fakultät), Universität Innsbruck, Austria.
Appl Environ Microbiol. 1997 Feb;63(2):753-6. doi: 10.1128/aem.63.2.753-756.1997.
A system for regulated heterologous gene expression in the filamentous fungus Penicillium chrysogenum was established. This is the first heterologous expression system to be developed for this organism. Expression of a recombinant fungal xylanase gene (xylp) and the cDNA for the human tear lipocalin (LCNI) was achieved by placing the encoding sequences under the control of the repressible acid phosphatase gene (phoA) promoter of P. chrysogenum. Secreted recombinant proteins were detected in the growth media of transformed P. chrysogenum cells by means of bioassays, zymogramography, and Western blotting. Levels of transcription and amounts of recombinant proteins secreted varied among transformants, mainly due to the copy number and the integration site of the expression vector on the fungal chromosome.
建立了一种用于在产黄青霉丝状真菌中调控异源基因表达的系统。这是为该生物体开发的首个异源表达系统。通过将编码序列置于产黄青霉可阻遏酸性磷酸酶基因（phoA）启动子的控制下，实现了重组真菌木聚糖酶基因（xylp）和人泪液脂钙蛋白（LCNI）cDNA的表达。通过生物测定、酶谱分析和蛋白质免疫印迹法在转化的产黄青霉细胞的生长培养基中检测到分泌的重组蛋白。转录水平和分泌的重组蛋白量在转化体之间有所不同，主要是由于表达载体在真菌染色体上的拷贝数和整合位点。
