Jeenes D J, Mackenzie D A, Archer D B
Department of Genetics and Microbiology, Institute of Food Research, Norwich Research Park, Colney, UK.
Transgenic Res. 1994 Sep;3(5):297-303. doi: 10.1007/BF01973589.
The relationship between heterologous gene copy number, mRNA and secreted protein yields has been studied in Aspergillus niger transformants containing either the hen egg-white lysozyme (HEWL) cDNA gene or a glucoamylase-HEWL gene fusion (incorporating the A. niger glaA gene). The results support a direct relationship between HEWL gene copy number, mRNA and secreted HEWL protein levels at low (< 25) copy numbers. High protein yields are associated with multiple copies of the recombinant gene at a single site. Fusion of the HEWL gene to the glucoamylase gene resulted in higher steady-state levels of heterologous mRNA. Transformants with the HEWL cDNA alone exhibited a ten-fold higher mRNA:protein ratio than transformants with the gene fusion indicating that post-transcriptional events significantly affect final secreted protein yields.
在含有鸡蛋清溶菌酶(HEWL)cDNA基因或葡糖淀粉酶-HEWL基因融合体(包含黑曲霉glaA基因)的黑曲霉转化体中,研究了异源基因拷贝数、mRNA和分泌蛋白产量之间的关系。结果表明,在低拷贝数(<25)时,HEWL基因拷贝数、mRNA和分泌的HEWL蛋白水平之间存在直接关系。高蛋白产量与重组基因在单个位点的多个拷贝相关。HEWL基因与葡糖淀粉酶基因的融合导致异源mRNA的稳态水平更高。仅含有HEWL cDNA的转化体比含有基因融合体的转化体表现出高十倍的mRNA:蛋白质比率,这表明转录后事件显著影响最终分泌蛋白的产量。