Gómez-Pardo E, Peñalva M A
Centro de Investigaciones Biológicas, Velázquez, Madrid, Spain.
Gene. 1990 Apr 30;89(1):109-15. doi: 10.1016/0378-1119(90)90212-a.
We have constructed a translational fusion between the isopenicillin-N-synthetase-encoding gene (IPNS) of Aspergillus nidulans and the lacZ gene of Escherichia coli. Recombinant strains carrying a single copy of the fusion integrated at the IPNS locus produced beta-galactosidase (beta Gal) during secondary metabolism. Integration of the fusion at the argB locus results in a situation in which the only 5'-flanking sequences of the IPNS gene upstream from the chimeric fused gene are those included in the transforming plasmid. Such a strain still expresses beta Gal activity during secondary metabolism, showing that a DNA fragment including sequences of the IPNS gene from nt -2000 to +35 (relative to the translation start codon) still contains sufficient information to drive expression of the fusion gene during secondary metabolism.
我们构建了构巢曲霉的异青霉素-N-合成酶编码基因(IPNS)与大肠杆菌lacZ基因之间的翻译融合体。携带融合体单拷贝并整合在IPNS基因座的重组菌株在次级代谢过程中产生β-半乳糖苷酶(βGal)。融合体整合到argB基因座会导致这样一种情况,即嵌合融合基因上游IPNS基因的唯一5'侧翼序列是转化质粒中包含的那些序列。这样的菌株在次级代谢过程中仍表现出βGal活性,表明一个包含IPNS基因从nt -2000到+35(相对于翻译起始密码子)序列的DNA片段仍然包含足够的信息来驱动融合基因在次级代谢过程中的表达。