Lenouvel François, van de Vondervoort Peter J I, Visser Jaap
Molecular Genetics of Industrial Micro-Organisms, Wageningen Agricultural University, Dreijenlaan 2, 6703 HA Wageningen, The Netherlands.
Curr Genet. 2002 Sep;41(6):425-32. doi: 10.1007/s00294-002-0320-0. Epub 2002 Aug 14.
We disrupted the Aspergillus niger gene argB, encoding ornithine transcarbamylase. Full characterisation of the argB deletion was performed by Southern blot analysis, growth tests and by means of mitotic recombination, complementation and transformation. The argB locus was found to be physically removed, thus creating an auxotrophic mutation. The latter can be supplemented by addition of arginine into the culture medium. The argB gene and its disruption do not correlate to the argI13 (formerly argB13) allele described. The delta argB is on chromosome I whereas argI13 is on V. In addition, the argI13 mutation can only be complemented by the A. nidulans argB gene, whereas the new argB deletion can be complemented by both the A. niger and A. nidulans argB genes. The delta argB strain has been used to generate several strains in a breeding programme and to study the expression of important genes, such as areA and kexB.
我们破坏了黑曲霉中编码鸟氨酸转氨甲酰酶的argB基因。通过Southern印迹分析、生长测试以及有丝分裂重组、互补和转化等手段,对argB缺失进行了全面表征。发现argB基因座已被物理移除,从而产生了一个营养缺陷型突变。后者可通过向培养基中添加精氨酸来补充。argB基因及其破坏与所描述的argI13(原argB13)等位基因不相关。ΔargB位于染色体I上,而argI13位于染色体V上。此外,argI13突变只能由构巢曲霉的argB基因互补,而新的argB缺失可由黑曲霉和构巢曲霉的argB基因互补。ΔargB菌株已被用于在育种计划中产生多个菌株,并用于研究重要基因如areA和kexB的表达。