RESUMO
The cloning and nucleotide sequence of the genes (idi, crtE, crtYB, crtl and crtS) controlling the astaxanthin biosynthesis pathway of the wild-type ATCC 24230 strain of Xanthophyllomyces dendrorhous in their genomic and cDNA version were obtained. The idi, crtE, crtYB, crtl and crtS genes were cloned, as fragments of 10.9, 11.5, 15.8, 5.9 and 4 kb respectively. The nucleotide sequence data analysis indicates that the idi, crtE, crtYB, crtl and crtS genes have 4, 8,4, 11, and 17 introns and 5, 9, 5, 12 and 18 exons respectively. In addition, a highly efficient site-directed mutagenesis system was developed by transformation by integration, followed by mitotic recombination (the double recombinant method). Heterozygote idi (idi+ / idi-::hph), crtE (crtE+ / crtE -::hph), crtYB (crtYB + / crtYB -::hph), crtI (crtI+ / crtI-::hph) and crtS (crtS +/crtS -::hph) and homozygote mutants crtYB (crtYB -::hph/crtYB -::hph), crtI (crtI -::hph/crtI -::hph) and crtS (crtS -::hph / crtS -::hph) were constructed. All the heterozygote mutants have a pale phenotype and produce less carotenoids than the wild-type strain. The genetic analysis of the crtYB, crtl and crtS loci in the wild-type, heterozygote, and homozygote give evidence of the diploid constitution of ATCC 24230 strains. In addition, the cloning of a truncated form of the crtYB that lacks 153 amino acids of the N-terminal region derived from alternatively spliced mRNA was obtained. Their heterologous expression in Escherichia coli carrying the carotenogenic cluster of Erwinia uredovora result in trans-complementation and give evidence of its functionality in this bacterium, maintaining its phytoene synthase activity but not the lycopene cyclase activity.
Assuntos
Basidiomycota/genética , Regulação Fúngica da Expressão Gênica/genética , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , DNA Complementar/genética , Genes Fúngicos/genética , Reação em Cadeia da Polimerase , RNA Fúngico/genética , Xantofilas/biossíntese , Xantofilas/genéticaRESUMO
Se realizó un estudio descriptivo retrospectivo, de corte transversal, en el área del Policlínico Docente "Wilfredo Pérez Pérez" del municipio San Miguel del Padrón, de Ciudad de La Habana, con el fin de explorar los conocimientos sobre algunos aspectos de la sexualidad, en un grupo de 100 adolescentes en edades entre 10 y 19 años y de uno y otro sexos, en el año 2001. Se apreció un elevado tanto por ciento de jóvenes que tenían conocimientos sobre la anatomía del aparato genital, la mejor edad para tener hijos y la anticoncepción. En general, no tenían conocimientos de todas las enfermedades de transmisión sexual, no así el método más eficaz para evitarlas. El grupo de edad de mayor relevancia lo constituyó el de 10 a 13 años por sus escasos conocimientos sobre sexualidad