RESUMEN
We report the first isolation and sequencing of genomic BAC clones containing the marsupial milk protein genes Whey Acidic Protein (WAP) and Early Lactation Protein (ELP). The stripe-faced dunnart WAPgene sequence contained five exons, the middle three of which code for the WAPmotifs and four disulphide core domains which characterize WAP. The dunnart ELPgene sequence contained three exons encoding a protein with a Kunitz motif common to serine protease inhibitors. Fluorescence in situ hybridization located the WAPgene to chromosome 1p in the stripe-faced dunnart, and the ELPgene to 2q. Northern blot analysis of lactating mammary tissue of the closely related fat-tailed dunnart has shown asynchronous expression of these milk protein genes. ELPwas expressed at only the earlier phase of lactation and WAPonly at the later phase of lactation, in contrast to beta-lactoglobulin (BLG) and alpha-lactalbumin (ALA) genes, which were expressed in both phases of lactation. This asynchronous expression during the lactation cycle in the fat-tailed dunnart is similar to other marsupials and it probably represents a pattern that is ancestral to Australian marsupials.
Asunto(s)
Marsupiales/fisiología , Proteínas de la Leche/genética , Animales , Aprotinina , Australia , Secuencia de Bases , Cromosomas , Exones , Femenino , Regulación de la Expresión Génica/fisiología , Lactancia/genética , Glándulas Mamarias Animales/metabolismo , Marsupiales/genética , Análisis de Secuencia de ADN , Proteína de Suero de LecheRESUMEN
Comparing globin genes and their flanking sequences across many species has allowed globin gene evolution to be reconstructed in great detail. Marsupial globin sequences have proved to be of exceptional significance. A previous finding of a beta(beta)-like omega(omega) gene in the alpha(alpha) cluster in the tammar wallaby suggested that the alpha and beta cluster evolved via genome duplication and loss rather than tandem duplication. To confirm and extend this important finding we isolated and sequenced BACs containing the alpha and beta loci from the distantly related Australian marsupial Sminthopsis macroura. We report that the alpha gene lies in the same BAC as the beta-like omega gene, implying that the alpha-omega juxtaposition is likely to be conserved in all marsupials. The LUC7L gene was found 3' of the S. macroura alpha locus, a gene order shared with humans but not mouse, chicken or fugu. Sequencing a BAC contig that contained the S. macroura beta globin and epsilon globin loci showed that the globin cluster is flanked by olfactory genes, demonstrating a gene arrangement conserved for over 180 MY. Analysis of the region 5' to the S. macroura epsilon (epsilon) globin gene revealed a region similar to the eutherian LCR, containing sequences and potential transcription factor binding sites with homology to eutherian hypersensitive sites 1 to 5. FISH mapping of BACs containing S. macroura alpha and beta globin genes located the beta globin cluster on chromosome 3q and the alpha locus close to the centromere on 1q, resolving contradictory map locations obtained by previous radioactive in situ hybridization.
Asunto(s)
Mapeo Cromosómico/métodos , Hemoglobinas/genética , Marsupiales/genética , Familia de Multigenes/genética , Análisis de Secuencia de ADN/métodos , Animales , Australia , Cromosomas Artificiales Bacterianos/genética , Secuencia Conservada/genética , Evolución Molecular , Galago/genética , Globinas/genética , Cabras/genética , Humanos , Ratones , Datos de Secuencia Molecular , Zarigüeyas/genética , Filogenia , ConejosRESUMEN
Reciprocal chromosome painting and G-banding were used to compare the karyotypes of three Australian marsupials (Sminthopsis crassicaudata, Macropus eugenii, Trichosurus vulpecula) and one South American marsupial (Monodelphis domestica). The results revealed only a limited number of rearrangements between these species and that the four karyotypes can be described as different combinations of fifteen conserved segments. Five chromosomes are totally conserved between M. domestica (pairs 1, 2, 5, 8 and the X) and the presumed 2n = 14 Australian ancestral karyotype, while M. domestica pairs 3 and 6 and 4 and 7 would have been involved in fusion/fission rearrangements. Chromosome comparisons are presented in a chromosome homology map. Although the species studied diverged 70 million years ago, the karyotype of Monodelphis domestica is highly conserved in relation to those of Australian marsupials.
Asunto(s)
Cromosomas/genética , Evolución Molecular , Marsupiales/genética , Animales , Australia , Bandeo Cromosómico , Pintura Cromosómica , Cariotipificación , Masculino , Homología de Secuencia de Ácido Nucleico , América del SurRESUMEN
A 2n = 14 karyotype is shared by some species in each of the marsupial orders in Australian and American superfamilies, suggesting that the ancestral marsupial chromosome complement was 2n = 14. We have used chromosome painting between distantly related marsupial species to discover whether genome arrangements in 2n = 14 species in two Australian orders support this hypothesis. Cross-species chromosome painting was used to investigate chromosome rearrangements between a macropodid species Macropus eugenii (2n = 16) and a wombat species in a different suborder (Lasiorhinus latifrons, 2n = 14), and a dasyurid species in a different order (Sminthopsis macroura, 2n = 14). We demonstrate that many chromosome regions are conserved between all three species, and deduce how the similar 2n = 14 karyotypes of species in the two orders are related to a common ancestral 2n = 14 karyotype.
Asunto(s)
Pintura Cromosómica , Marsupiales/genética , Animales , Secuencia de Bases , Bandeo Cromosómico , Cartilla de ADN , Cariotipificación , Especificidad de la Especie , Cromosoma X , Cromosoma YRESUMEN
In order to deduce the ancestral genome arrangement in the karyotypically diverse marsupial family Macropodidae, and to assess chromosome change in this family, chromosome-specific paints from the tammar wallaby (2n = 16) were hybridized to metaphase spreads from the two species proposed to represent the 2n = 22 ancestral karyotype, as well as species with derived 2n = 20 and 2n = 14 karyotypes. Identical patterns were observed in the two 2n = 22 species, from which the rearrangements to form the three derived karyotypes may be easily deduced to be 1, 3 and 4 different fusions, respectively. The identical Thylogale and Dorcopsis genomes may both be used to represent the pleisiomorphic macropodid chromosome complement. Variation in the X chromosome was also investigated by hybridizing an X-Y shared tammar wallaby 12-kb repeat element to chromosomes from the other four macropodid species, finding that it hybridized only to the most closely related species, and therefore is of recent origin.
