Assuntos
Clonagem Molecular , Desoxirribonucleases de Sítio Específico do Tipo II/metabolismo , Vetores Genéticos , Reação em Cadeia da Polimerase , Sequência de Bases , Meios de Cultura , Desoxirribonucleases de Sítio Específico do Tipo II/normas , Escherichia coli/genética , Técnicas de Transferência de Genes , Dados de Sequência Molecular , Oligonucleotídeos/genética , Oligonucleotídeos/metabolismoRESUMO
The organization of the gene encoding potato UDP-glucose pyrophosphorylase, one of the key enzymes of carbohydrate metabolic pathway is presented. The gene cloned from cultivar (cv.) Lemhi consists of a 6.6-kb structural and a 1-kb regulatory region. The structural region contains 20 exons and 19 introns. The coding sequence with exception of three bases is identical with the UGPase cDNA previously cloned from Danshaku-Imo cv. [Katsube et al. (1990) UDP-Glucose pyrophosphorylase from potato tuber: cDNA cloning and sequencing. J. Biochem. 108, 321-326]. The largest intron contains a tandem repeat consisting of 50 nt core units. A putative polyadenylation site is situated 79 bp downstream of the translation stop codon. A transcription start point (tsp) and a putative TATA-box were located 84 bp and 141 bp upstream of the translation start, respectively. The regulatory region contained general enhancer, suppressor, and regions responsible for tissue specificity of UGPase expression.
Assuntos
Solanum tuberosum/enzimologia , Solanum tuberosum/genética , Transcrição Gênica , UTP-Glucose-1-Fosfato Uridililtransferase/biossíntese , UTP-Glucose-1-Fosfato Uridililtransferase/genética , Sequência de Bases , Clonagem Molecular , Éxons , Íntrons , Dados de Sequência Molecular , Biossíntese de Proteínas , Proteínas Recombinantes/biossíntese , Sequências Reguladoras de Ácido Nucleico , Mapeamento por RestriçãoRESUMO
A 747-bp tandem repeat element from the genome of the fungus Plasmopara halstedii, the causal agent of downy mildew of sunflower, was cloned and analyzed. The clone can be used as a probe to distinguish races of the pathogen. Sequence analysis of a copy of this element revealed the presence of 103 direct repeats of 6 bp or greater and two potential ORFs. This tandemly repeated element consists of four subunits that may have evolved as a result of several unequal crossing-over events.
