RESUMO
A second gene (rp-L21) copy, clone g34, coding for ribosomal (r-) protein L21, was isolated from the pathogenic (P) strain HM-1:IMSS cl6 of the intestinal parasite Entamoeba histolytica (Eh). The gene was compared to the previously isolated copy, gLE3 [Petter et al., Mol. Biochem. Parasitol. 56 (1992) 329-334], with respect to its primary structure, mRNA levels and binding to the r-complex during translation. Unlike the gLE3 gene copy [Petter et al., Mol. Biochem. Parasitol. 56 (1992) 329-334], g34 was found not to be physically connected to an actin gene copy. Homologous copies of the two rp-L21 genes were also characterized from the nonpathogenic (NP) strain SAW1734R clAR, as well as from its P derivative. Sequence comparison of the coding regions of the two rp-L21 revealed almost full identity. Significant differences were found, however, within their 3' and 5' flanking regions. Using the 3' rapid amplification of cDNA ends (3' RACE) method [Frohman et al., Proc. Natl. Acad. Sci. USA 85 (1988) 8998-9002], as well as Northern and slot blot hybridizations, it was demonstrated that both rp-L21 mRNAs are found in similar amounts. However, as was shown by differential hybridization, the relative binding of each transcript to the r-complex varied somewhat between P and NP strains. This finding suggests that the control of expression of rp-L21 in Eh may involve regulation at the post-transcriptional level.
Assuntos
Entamoeba histolytica/genética , Proteínas de Protozoários/genética , RNA de Protozoário , Proteínas Ribossômicas/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , DNA de Protozoário , Entamoeba histolytica/patogenicidade , Genes de Protozoários , Dados de Sequência Molecular , RNA Mensageiro/metabolismo , RNA de Protozoário/metabolismo , Alinhamento de SequênciaRESUMO
DNA hybridization with oligonucleotide probes is a powerful technique to study population genetics and structures. The use of probes which recognize ubiquitously interspersed DNA sequences has a distinct advantage over other techniques (e.g. the analysis of patterns of restriction fragment length polymorphism) in that many independent loci can be detected simultaneously. In this communication, we investigated the use of a trinucleotide repetitive DNA oligonucleotide, poly(GTG)5, in Southern blot analysis of Salmonella serotypes and Shigella species. The strains in this study were isolated over several years from widely disparate geographic locations and can therefore be considered to represent the structure of part of the natural populations of these organisms. In most of the Salmonella serotypes, the poly(GTG)5-associated profile (GTG profile) phenotypes appeared to be clonally stable; in cases where only one isolate of a serotype was tested, the GTG profile was distinct from the others. On the other hand, when GTG profile analysis was applied to Shigella strains, each of the 12 isolates, belonging to the four Shigella species, produced a unique pattern phenotype of both the chromosome and plasmid DNA.
Assuntos
DNA Bacteriano/análise , Hibridização Genética/genética , Oligonucleotídeos/genética , Salmonella/genética , Shigella/genética , Autorradiografia , Técnicas de Tipagem Bacteriana , Southern Blotting , Técnicas In Vitro , Fenótipo , Plasmídeos/genéticaRESUMO
Several lines of evidence indicate a relatively low genetic heterogeneity in the natural Salmonella typhi population. However, some S. typhi isolates found in Indonesia express, instead of the usual fliC-d flagellin gene, a different flagellar gene fliC-j. In addition, Indonesian strains may have a second flagellar antigen fliC-z66. We have previously suggested, on the basis of the flagellar antigen constitution, that S. typhi evolved in an isolated human population in Indonesia. In order to test this hypothesis, we have gathered S. typhi isolates from around the world and tested the genetic heterogeneity among them. In general, polymorphism was greater in isolates from the Far East, as was indicated by Southern hybridizations with rDNA and fliC DNA probes. Gene fliC-j was not found in S. typhi isolates, other than those from Indonesia. However, the one-clone origin of S. typhi was indicated by a common DNA fingerprint pattern and by the occurrence, in the 5' end region of the fliC gene, of 10 scattered nucleotides that differ from the corresponding 10 nucleotides in other fliC alleles studied. These nucleotides were present in all isolates tested but did not change the amino acid sequence of the flagellin polypeptide.
