Harmsen J A, Kusters-van Someren M A, Visser J
Department of Genetics, Agricultural University, Wageningen, The Netherlands.
Curr Genet. 1990 Aug;18(2):161-6. doi: 10.1007/BF00312604.
Using the previously cloned Aspergillus niger N756 pectin lyase D gene as a probe, the corresponding pelD gene has been isolated from a genomic library of the laboratory strain A. niger N400. This gene encodes PLD, previously described as PLI, which is one of the two major pectin lyases isolated from the commercial pectinase preparation Ultrazym. Heterologous hybridization of the A. niger N400 genomic library with the pelD gene led to the isolation of another five genes: pelA, B, C, E, and F. These genes differ in their hybridization patterns with probes containing either the entire pelD gene, or 5' or 3' parts thereof. By partial sequencing, and expression in an A. niger transformant containing multiple copies of the pelA gene, we show that this gene, which hybridizes strongest with the pelD gene, encodes the other major pectin lyase from Ultrazym, PLII.
以先前克隆的黑曲霉N756果胶裂解酶D基因作为探针,从实验室菌株黑曲霉N400的基因组文库中分离出了相应的pelD基因。该基因编码PLD,以前称为PLI,它是从商业果胶酶制剂Ultrazym中分离出的两种主要果胶裂解酶之一。用pelD基因对黑曲霉N400基因组文库进行异源杂交,又分离出另外五个基因:pelA、B、C、E和F。这些基因与包含整个pelD基因或其5'或3'部分的探针的杂交模式不同。通过部分测序以及在含有多个拷贝pelA基因的黑曲霉转化体中的表达,我们表明,这个与pelD基因杂交最强的基因编码了Ultrazym中的另一种主要果胶裂解酶PLII。