Feng G H, Chu F S, Leonard T J
Department of Botany, University of Wisconsin, Madison 53706.
Appl Environ Microbiol. 1992 Feb;58(2):455-60. doi: 10.1128/aem.58.2.455-460.1992.
A differential hybridization strategy was used to clone genes associated with aflatoxin biosynthesis. A genomic library, formed between nuclear DNA and the pUC19 plasmid, was screened with three different cDNA probes by the colony hybridization procedure. Nineteen clones were selected; all were positively correlated with and presumably enriched with genes associated with aflatoxin production. Some of these clones were further characterized by using them as probes in Northern (RNA blot) hybridizations. Five clones hybridized strongly with some polyadenylated RNAs formed during the transition to or during idiophase when aflatoxin was produced. However, little or no corresponding hybridization occurred with polyadenylated RNAs formed in early and mid-log growth phase. Two of the clones were further used as probes to hybridize with polyadenylated RNAs formed under aflatoxin-permissive and nonpermissive temperatures. Hybridization occurred with RNA species formed under the permissive temperature only.
采用差异杂交策略克隆与黄曲霉毒素生物合成相关的基因。通过菌落杂交程序，用三种不同的cDNA探针筛选了由核DNA与pUC19质粒构建的基因组文库。挑选出19个克隆；所有克隆均与黄曲霉毒素产生相关的基因呈正相关且可能富含这些基因。其中一些克隆通过在Northern（RNA印迹）杂交中用作探针进行了进一步表征。五个克隆与黄曲霉毒素产生时过渡到分化期或分化期形成的一些多聚腺苷酸化RNA强烈杂交。然而，在对数生长早期和中期形成的多聚腺苷酸化RNA几乎没有或没有发生相应的杂交。其中两个克隆进一步用作探针，与在黄曲霉毒素允许和非允许温度下形成的多聚腺苷酸化RNA杂交。仅与允许温度下形成的RNA种类发生杂交。
