Wan Y, Liu H, Li C, Schmidhauser T J
University of Colorado Health Sciences Center, Denver 80262, USA.
Fungal Genet Biol. 1997 Jun;21(3):329-36. doi: 10.1006/fgbi.1997.0988.
Two chromosome walks covering 420 and 110 kb on the left arm of linkage group VI (LGVIL) of Neurospora crassa were purscrooued with the goal of cloning carotenogenic loci. Complementation analysis with clones isolated in the 420-kb walk allowed identification of the yellow-1 (ylo-1) gene which is essential for Neurospora carotenogenesis. We have physically located a second gene, unknown-13 (un-13), between the cross-pathway control-1, (cpc-1) and ylo-1 loci. Cloning of a second potential carotenogenic locus, vivid (vvd), from our walks was attempted using screening of Northern blots with radiolabeled DNA fragments from walk clones to identify gene transcripts. The radiolabeled DNA fragments were used to clone complementary DNA isolates representing an additional four genes in the two walks.
为了克隆类胡萝卜素生成基因座，对粗糙脉孢菌VI连锁群（LGVIL）左臂上覆盖420和110 kb的两个染色体步移进行了筛选。用在420 kb步移中分离的克隆进行互补分析，从而鉴定出了黄色-1（ylo-1）基因，该基因对粗糙脉孢菌的类胡萝卜素生成至关重要。我们已将第二个基因，未知-13（un-13），定位在交叉途径控制-1（cpc-1）和ylo-1基因座之间。尝试从我们的步移中克隆第二个潜在的类胡萝卜素生成基因座，即生动（vvd）基因座，方法是用来自步移克隆的放射性标记DNA片段筛选Northern印迹，以鉴定基因转录本。放射性标记的DNA片段被用于克隆代表这两个步移中另外四个基因的互补DNA分离物。
