Your browser doesn't support javascript.
loading
Bulk segregant analysis followed by high-throughput sequencing reveals the Neurospora cell cycle gene, ndc-1, to be allelic with the gene for ornithine decarboxylase, spe-1.
Pomraning, Kyle R; Smith, Kristina M; Freitag, Michael.
Afiliación
  • Pomraning KR; Department of Biochemistry and Biophysics, 2011 ALS Bldg., Oregon State University, Corvallis, OR 97331-7305, USA.
Eukaryot Cell ; 10(6): 724-33, 2011 Jun.
Article en En | MEDLINE | ID: mdl-21515825
ABSTRACT
With the advent of high-throughput DNA sequencing, it is now straightforward and inexpensive to generate high-density small nucleotide polymorphism (SNP) maps. Here we combined high-throughput sequencing with bulk segregant analysis to expedite mutation mapping. The general map location of a mutation can be identified by a single backcross to a strain enriched in SNPs compared to a standard wild-type strain. Bulk segregant analysis simultaneously increases the likelihood of determining the precise nature of the mutation. We present here a high-density SNP map between Neurospora crassa Mauriceville-1-c (FGSC2225) and OR74A (FGSC2489), the strains most typically used by Neurospora researchers to carry out mapping crosses. We further have demonstrated the utility of the Mauriceville sequence and our approach by mapping the mutation responsible for the only existing temperature-sensitive (ts) cell cycle mutation in Neurospora, nuclear division cycle-1 (ndc-1). The single T-to-C point mutation maps to the gene encoding ornithine decarboxylase (ODC), spe-1 (NCU01271), and changes a Phe to a Ser residue within a highly conserved motif next to the catalytic site of the enzyme. By growth on spermidine and complementation with a wild-type spe-1 gene, we showed that the defect in spe-1 is responsible for the ts ndc-1 mutation. Based on our results, we propose changing ndc-1 to spe-1(ndc), which reflects that this mutation results in an ODC with a specific nuclear division defect.
Asunto(s)

Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Ornitina Descarboxilasa / Proteínas Fúngicas / Análisis de Secuencia de ADN / Proteínas de Ciclo Celular / Neurospora crassa Idioma: En Revista: Eukaryot Cell Asunto de la revista: BIOLOGIA MOLECULAR Año: 2011 Tipo del documento: Article País de afiliación: Estados Unidos

Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Ornitina Descarboxilasa / Proteínas Fúngicas / Análisis de Secuencia de ADN / Proteínas de Ciclo Celular / Neurospora crassa Idioma: En Revista: Eukaryot Cell Asunto de la revista: BIOLOGIA MOLECULAR Año: 2011 Tipo del documento: Article País de afiliación: Estados Unidos