Dutta S K, Chaudhuri R K
Mol Gen Genet. 1978 Aug 4;164(1):45-50. doi: 10.1007/BF00267597.
Transcriptional properties of an "albino" mutant, a "slime" mutant and a wild type strain of Neurospora crassa have been compared by quantitative estimates of DNA:RNA hybridization. The RNAs isolated from these strains were incubated with 32P-DNA of the wild type strain. The DNA:RNA hybrids were isolated by chromatography on hydroxyapatite. 29--34% of the wild type 32P-DNA hybridized with RNA from all of the strains used. The DNA sequences which gave a primary reaction with wild type RNA (tr-DNA) were recovered and again reacted with different RNAs. The tr-DNA sequences reacted to the extent of 92% with the wild type RNA, but only 78% with slime mutant RNA and 85% with the RNA isolated from the albino mutant. The use of tr-DNA sequences thus isolated, proved to have increased the specificity of the hybridization reactions, these were useful indicators for identification of transcriptional differences among wild type and mutant strains of N. crassa.
通过对DNA:RNA杂交的定量估计,比较了粗糙脉孢菌“白化”突变体、“黏液”突变体和野生型菌株的转录特性。从这些菌株中分离出的RNA与野生型菌株的32P-DNA一起孵育。DNA:RNA杂交体通过羟基磷灰石层析分离。29%-34%的野生型32P-DNA与所用所有菌株来源的RNA杂交。回收了与野生型RNA发生初次反应的DNA序列(tr-DNA),并再次与不同的RNA反应。tr-DNA序列与野生型RNA的反应程度为92%,但与黏液突变体RNA的反应程度仅为78%,与从白化突变体中分离出的RNA的反应程度为85%。如此分离得到的tr-DNA序列被证明提高了杂交反应的特异性,这些是鉴定粗糙脉孢菌野生型和突变体菌株转录差异的有用指标。