RESUMO
Growing evidence suggests that the gut microbiome (GM) plays a critical role in health and disease. However, the contribution of GM to psychiatric disorders, especially anxiety, remains unclear. We used the Collaborative Cross (CC) mouse population-based model to identify anxiety associated host genetic and GM factors. Anxiety-like behavior of 445 mice across 30 CC strains was measured using the light/dark box assay and documented by video. A custom tracking system was developed to quantify seven anxiety-related phenotypes based on video. Mice were assigned to a low or high anxiety group by consensus clustering using seven anxiety-related phenotypes. Genome-wide association analysis (GWAS) identified 141 genes (264 SNPs) significantly enriched for anxiety and depression related functions. In the same CC cohort, we measured GM composition and identified five families that differ between high and low anxiety mice. Anxiety level was predicted with 79% accuracy and an AUC of 0.81. Mediation analyses revealed that the genetic contribution to anxiety was partially mediated by the GM. Our findings indicate that GM partially mediates and coordinates the effects of genetics on anxiety.
Assuntos
Ansiedade/genética , Ansiedade/microbiologia , Comportamento Animal , Microbioma Gastrointestinal , Animais , Camundongos de Cruzamento Colaborativo , CamundongosRESUMO
We constructed a bacterial artificial chromosome (BAC)-based physical map of chromosomes 2 and 3 of Drosophila melanogaster, which constitute 81% of the genome. Sequence tagged site (STS) content, restriction fingerprinting, and polytene chromosome in situ hybridization approaches were integrated to produce a map spanning the euchromatin. Three of five remaining gaps are in repeat-rich regions near the centromeres. A tiling path of clones spanning this map and STS maps of chromosomes X and 4 was sequenced to low coverage; the maps and tiling path sequence were used to support and verify the whole-genome sequence assembly, and tiling path BACs were used as templates in sequence finishing.
Assuntos
Mapeamento de Sequências Contíguas , Drosophila melanogaster/genética , Genoma , Animais , Centrômero/genética , Cromatina/genética , Cromossomos Bacterianos/genética , Clonagem Molecular , Impressões Digitais de DNA , Eucromatina , Biblioteca Gênica , Genes de Insetos , Marcadores Genéticos , Vetores Genéticos , Hibridização In Situ , Sequências Repetitivas de Ácido Nucleico , Mapeamento por Restrição , Análise de Sequência de DNA , Sitios de Sequências Rotuladas , Telômero/genéticaRESUMO
The fly Drosophila melanogaster is one of the most intensively studied organisms in biology and serves as a model system for the investigation of many developmental and cellular processes common to higher eukaryotes, including humans. We have determined the nucleotide sequence of nearly all of the approximately 120-megabase euchromatic portion of the Drosophila genome using a whole-genome shotgun sequencing strategy supported by extensive clone-based sequence and a high-quality bacterial artificial chromosome physical map. Efforts are under way to close the remaining gaps; however, the sequence is of sufficient accuracy and contiguity to be declared substantially complete and to support an initial analysis of genome structure and preliminary gene annotation and interpretation. The genome encodes approximately 13,600 genes, somewhat fewer than the smaller Caenorhabditis elegans genome, but with comparable functional diversity.
Assuntos
Drosophila melanogaster/genética , Genoma , Análise de Sequência de DNA , Animais , Transporte Biológico/genética , Cromatina/genética , Clonagem Molecular , Biologia Computacional , Mapeamento de Sequências Contíguas , Sistema Enzimático do Citocromo P-450/genética , Reparo do DNA/genética , Replicação do DNA/genética , Drosophila melanogaster/metabolismo , Eucromatina , Biblioteca Gênica , Genes de Insetos , Heterocromatina/genética , Proteínas de Insetos/química , Proteínas de Insetos/genética , Proteínas de Insetos/fisiologia , Proteínas Nucleares/genética , Biossíntese de Proteínas , Transcrição GênicaRESUMO
A comparative analysis of the genomes of Drosophila melanogaster, Caenorhabditis elegans, and Saccharomyces cerevisiae-and the proteins they are predicted to encode-was undertaken in the context of cellular, developmental, and evolutionary processes. The nonredundant protein sets of flies and worms are similar in size and are only twice that of yeast, but different gene families are expanded in each genome, and the multidomain proteins and signaling pathways of the fly and worm are far more complex than those of yeast. The fly has orthologs to 177 of the 289 human disease genes examined and provides the foundation for rapid analysis of some of the basic processes involved in human disease.
Assuntos
Caenorhabditis elegans/genética , Drosophila melanogaster/genética , Genoma , Proteoma , Saccharomyces cerevisiae/genética , Animais , Apoptose/genética , Evolução Biológica , Caenorhabditis elegans/química , Caenorhabditis elegans/fisiologia , Adesão Celular/genética , Ciclo Celular/genética , Drosophila melanogaster/química , Drosophila melanogaster/fisiologia , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Genes Duplicados , Doenças Genéticas Inatas/genética , Genética Médica , Proteínas de Helminto/química , Proteínas de Helminto/genética , Humanos , Imunidade/genética , Proteínas de Insetos/química , Proteínas de Insetos/genética , Família Multigênica , Neoplasias/genética , Estrutura Terciária de Proteína , Saccharomyces cerevisiae/química , Saccharomyces cerevisiae/fisiologia , Transdução de Sinais/genéticaRESUMO
The past year has been a spectacular one for Drosophila research. The sequencing and annotation of the Drosophila melanogaster genome has allowed a comprehensive analysis of the first three eukaryotes to be sequenced-yeast, worm and fly-including an analysis of the fly's influences as a model for the study of human disease. This year has also seen the initiation of a full-length cDNA sequencing project and the first analysis of Drosophila development using high-density DNA microarrays containing several thousand Drosophila genes. For the first time homologous recombination has been demonstrated in flies and targeted gene disruptions may not be far off.
Assuntos
Drosophila melanogaster/genética , Genes de Insetos , Genoma , Animais , Expressão Gênica , Predisposição Genética para Doença , Humanos , Proteínas de Insetos/genética , Proteínas de Insetos/fisiologia , Recombinação GenéticaRESUMO
To decipher and manipulate the 14 000 identified Drosophila genes, there is a need to inject a large number of embryos with transgenes. We have developed an automated instrument for high throughput injection of Drosophila embryos. It was built on an inverted microscope, equipped with a motorized xy stage, autofocus, a charge coupled device camera, and an injection needle mounted on a high speed vertical stage. A novel, micromachined embryo alignment device was developed to facilitate the arrangement of a large number of eggs. The control system included intelligent and dynamic imaging and analysis software and an embryo injection algorithm imitating a human operator. Once the injection needle and embryo slide are loaded, the software automatically images and characterizes each embryo and subsequently injects DNA into all suitable embryos. The ability to program needle flushing and monitor needle status after each injection ensures reliable delivery of biomaterials. Using this instrument, we performed a set of transformation injection experiments. The robot achieved injection speeds and transformation efficiencies comparable to those of a skilled human injector. Because it can be programed to allow injection at various locations in the embryo, such as the anterior pole or along the dorsal or ventral axes, this system is also suitable for injection of general biochemicals, including drugs and RNAi.
Assuntos
DNA/administração & dosagem , Drosophila/embriologia , Drosophila/genética , Injeções/instrumentação , Robótica/instrumentação , Transfecção/instrumentação , Animais , Animais Geneticamente Modificados , DNA/genética , Desenho de Equipamento , Análise de Falha de Equipamento , Injeções/métodos , Reprodutibilidade dos Testes , Robótica/métodos , Sensibilidade e Especificidade , Manejo de Espécimes/instrumentação , Manejo de Espécimes/métodos , Transfecção/métodosRESUMO
BACKGROUND: In the fruitfly Drosophila melanogaster, segment identity is specified by the homoeotic selector genes of the bithorax and Antennapedia complexes. The functions of these genes in the segmental specification of the Drosophila ectoderm have been well studied, but their roles in muscle development have been relatively poorly investigated. Recent experiments have strongly suggested that homeotic selector genes are directly involved in one aspect of mesodermal patterning during Drosophila embryogenesis. But muscle development is a complex process, requiring for its completion the correct positioning of the epidermis, the nervous system and the developing muscles in a segment-specific manner. Many aspects of homeotic selector gene function in this process remain to be understood. RESULTS: In flies that are homozygous for three mutant alleles (anterobithorax, bithorax3, postbithorax) of the Ultrabithorax gene, the third thoracic segment (T3) is transformed towards the second (T2). The adults have two pairs of wings, but the homeotically transformed T3 (HT3) has only rudimentary indirect flight muscles. We used the 'four-winged' fly to study the role of homeotic selector genes in the development of the indirect flight muscles, which we classify into four 'events'. First, the determination of the segment-specific pattern of myoblasts in the larval thorax; second, the specific pattern of migration of myoblasts during metamorphosis; third, the fusion of myoblasts to form adult indirect flight muscles and fourth, the development of the branching pattern of adult motor innervation. Our study shows that the segmental identity of the epidermis determines the segment-specific pattern and number of myoblasts on the larval discs, and the pattern of their migration during metamorphosis. The segmental identity of the mesoderm, however, is crucial for the fusion of myoblasts to form indirect flight muscles, and also influences the branching pattern of innervation of indirect flight muscles. CONCLUSIONS: Segmental information expressed in the ectoderm, and the autonomous function of homeotic selector genes in the mesoderm, are both required for the complete development of indirect flight muscles.
Assuntos
Proteínas de Drosophila , Drosophila melanogaster/crescimento & desenvolvimento , Drosophila melanogaster/genética , Genes Homeobox , Genes de Insetos , Proteínas de Homeodomínio , Desenvolvimento Muscular , Fatores de Transcrição , Alelos , Animais , Contagem de Células , Movimento Celular/genética , Proteínas de Ligação a DNA/genética , Drosophila melanogaster/citologia , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Larva/citologia , Larva/crescimento & desenvolvimento , Masculino , Metamorfose Biológica , Neurônios Motores/citologia , Músculos/citologia , Músculos/inervação , Mutação , Asas de Animais/citologia , Asas de Animais/crescimento & desenvolvimentoRESUMO
DNAs that contain specific yeast chromosomal sequences called ARSs transform Saccharomyces cerevisiae at high frequency and can replicate extrachromosomally as plasmids when introduced into S. cerevisiae by transformation. To determine the boundaries of the minimal sequences required for autonomous replication in S. cerevisiae, we have carried out in vitro mutagenesis of the first chromosomal ARS described, ARS1. Rather than identifying a distinct and continuous segment that mediates the ARS+ phenotype, we find three different functional domains within ARS1. We define domain A as the 11-base-pair (bp) sequence that is also found at most other ARS regions. It is necessary but not sufficient for high-frequency transformation. Domain B, which cannot mediate high-frequency transformation, or replicate by itself, is required for efficient, stable replication of plasmids containing domain A. Domain B, as we define it, is continuous with domain A in ARS1, but insertions of 4 bp between the two do not affect replication. The extent of domain B has an upper limit of 109 bp and a lower limit of 46 bp in size. There is no obvious sequence homology between domain B of ARS1 and any other ARS sequence. Finally, domain C is defined on the basis of our deletions as at least 200 bp flanking domain A on the opposite side from domain B and is also required for the stability of domain A in S. cerevisiae. The effect of deletions of domain C can be observed only in the absence of domain B, at least by the assays used in the current study, and the significance of this finding is discussed.
Assuntos
DNA Fúngico/genética , Mutação , Replicon , Saccharomyces cerevisiae/genética , Sequência de Bases , Deleção Cromossômica , Mapeamento Cromossômico , Replicação do DNA , Genes Fúngicos , Transformação GenéticaRESUMO
An enzyme system prepared from Saccharomyces cerevisiae carries out the replication of exogenous yeast plasmid DNA. Replication in vitro mimics that in vivo in that DNA synthesis in extracts of strain cdc8, a temperature-sensitive DNA replication mutant, is thermolabile relative to the wild-type, and in that aphidicolin inhibits replication in vitro. Furthermore, only plasmids containing a functional yeast replicator, ARS, initiate replication at a specific site in vitro. Analysis of replicative intermediates shows that plasmid YRp7, which contains the chromosomal replicator ARS1, initiates bidirectional replication in a 100 bp region within the sequence required for autonomous replication in vivo. Plasmids containing ARS2, another chromosomal replicator, and the ARS region of the endogenous yeast plasmid 2 microns circle give similar results, suggesting that ARS sequences are specific origins of chromosomal replication. Used in conjunction with deletion mapping, the in vitro system allows definition of the minimal sequences required for the initiation of replication.
Assuntos
Replicação do DNA , Saccharomyces cerevisiae/genética , Afidicolina , Ciclo Celular , Núcleo Celular/metabolismo , Clonagem Molecular , DNA Polimerase II/antagonistas & inibidores , Replicação do DNA/efeitos dos fármacos , Enzimas de Restrição do DNA , Diterpenos/farmacologia , Cinética , Mutação , Plasmídeos , Saccharomyces cerevisiae/efeitos dos fármacos , Moldes GenéticosRESUMO
Transabdominal (Tab), a dominant mutation in the Bithorax Complex (BX-C) of Drosophila, creates a sexually dimorphic pattern of segmental transformation that has complete penetrance and expressivity. Specific regions within the notum of the second thoracic segment (T2) are transformed into abdominal-like cuticle; thus, the Tab/ + notum has sets of short stripes that are black in males and only bordered with black in females. Also, Tab/ + abdominal tergites, A1-A6, inclusive, have small patches of A7-like tergite cuticle. Tab is inseparable from an 89E/90D inversion, whose DNA breakpoint in 89E is at +188 kb in the infra-abdominal-8 (iab-8) region of the BX-C. When probed with a pupal cDNA from the iab-7 region, labeling above background was not detected in wild-type wing discs but was detected in, and confined to, the notal region of Tab/ + wing discs. The Tab/ + phenotype is assumed to result from cis-overexpression of iab-7 in localized regions of segments T2-A6, inclusive.
Assuntos
Drosophila/genética , Genes Dominantes , Mutação , Animais , Cruzamentos Genéticos , DNA/genética , Drosophila/anatomia & histologia , Feminino , Genes Recessivos , Masculino , Hibridização de Ácido Nucleico , Fenótipo , TóraxRESUMO
Transabdominal (Tab) is a dominant gain-of-function mutation that results in islands of sexually dimorphic abdominal cuticle in the dorsal thorax of the adult fly. This phenotype has complete penetrance and constant expressivity, and we show that it results from ectopic expression of ABD-BII, one of two proteins derived from the Abdominal B (Abd-B) domain of the bithorax complex (BX-C) and one that is normally expressed only in terminal portions of the abdomen. In Tab/+ animals ABD-BII is ectopically expressed in the relevant imaginal "wing" disc as three islands of cells whose location on the fate map corresponds to the three islands of transformed cuticle in each half of the adult thorax. Tab is associated with an inseparable inversion bringing sequences in 90E next to sequences in the transcription unit encoding ABD-BII in 89E. That 90E sequences drive ectopic expression of ABD-BII is indicated by our finding that such sequences in a P-element transformant express the reporter gene's product (beta-galactosidase) in the same three islands of wing disc cells. On morphological grounds, the transformed islands in the adult thorax correspond to subsets of muscle attachment cells. Ectopic expression of a homeodomain protein thus creates a unique and invariant pattern of sexual dimorphism.
Assuntos
Drosophila/fisiologia , Genes Homeobox , Mutação , Caracteres Sexuais , Animais , Elementos Antissenso (Genética) , Sequência de Bases , Elementos de DNA Transponíveis , Proteínas de Ligação a DNA/genética , Drosophila/genética , Embrião não Mamífero/fisiologia , Feminino , Hibridização In Situ , Masculino , Dados de Sequência Molecular , Proteínas Recombinantes/metabolismo , Mapeamento por Restrição , Tórax , Transcrição Gênica , Asas de Animais/anatomia & histologia , beta-Galactosidase/genética , beta-Galactosidase/metabolismoRESUMO
In Drosophila the Abdominal-B (Abd-B) domain of the bithorax complex specifies the identities of several posterior abdominal segments, comprises homeo-protein-coding regions and cis-regulatory regions, and extends from infra-abdominal-5 (iab-5) to iab-8, inclusive. Mutations that eliminate the Abd-B domain act as late embryonic lethals and result in transformations of posterior abdominal segments toward more anterior ones. The Abd-B domain gives rise to a minimum of five homeo-box-containing transcripts, 7.8, 4.7, 4.3, 3.7, and 3.3 kb in length. We examined the structure of the Abd-B domain by sequencing two Abd-B cDNA clones derived from the 4.3- and the 4.7-kb transcripts and the corresponding genomic DNA. The domain spans approximately 100 kb and contains at least eight exons. The 4.7- and 4.3-kb transcripts contain an open reading frame capable of encoding a 54-kD protein. A portion of the deduced protein-coding sequence common to all of the Abd-B transcripts was cloned into an expression vector. The resultant fusion protein then was used to derive a monoclonal antibody specific to Abd-B. By use of that antibody, we identified two embryonic Abd-B proteins, 54 and 36 kD and determined the sum of their segmental distribution by immunohistochemical analysis of whole-mounted embryos and immunofluorescent analysis of dissected embryonic nervous systems. The proteins are distributed in the fourth to the ninth abdominal segments [parasegments (PS) 10-15] inclusive. Embryos homozygous for Polycomb (Pc) show labeling over almost the entire embryo, whereas embryos deficient for the Abd-B domain show no detectable labeling.
Assuntos
Proteínas de Drosophila , Drosophila melanogaster/genética , Genes Homeobox , Proteínas de Homeodomínio , Hormônios de Inseto/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , DNA/genética , Drosophila melanogaster/embriologia , Embrião não Mamífero/análise , Desenvolvimento Embrionário e Fetal , Regulação da Expressão Gênica , Humanos , Hormônios de Inseto/análise , Camundongos , Dados de Sequência Molecular , RNA Mensageiro/genética , Proteínas Recombinantes de Fusão/biossíntese , Homologia de Sequência do Ácido NucleicoRESUMO
Using markers that are expressed at muscle attachment sites, we have examined the early pupal development (first 36 hr) of Indirect Flight Muscle (IFM) attachments in the fruit fly Drosophila melanogaster. Expression of the Drosophila homologs of vertebrate integrins, the Position-Specific (PS) antigens, is known to differentially mark epidermal (PS1alpha) and muscle (PS2alpha) components of the developing IFM attachment sites. During myogenesis, PS2alpha is detected transiently in imaginal myoblasts that fuse with persistent larval muscles to give rise to the Dorsal Longitudinal Muscles (DLMs), but not in myoblasts that fuse de novo to give rise to the Dorso Ventral Muscles. The integrins are not expressed at attachment sites when the muscle fibers first make their appearance (12-20 hr). Following muscle-epidermal contact, PS1 and PS2 are detected at muscle attachment sites. PS1 expression is at the muscle ends and also in the long epidermal processes that connect the developing muscle fibers to their sites of attachment in the epidermis, while PS2 expression is restricted to the muscle ends. Epidermal cells that will contribute to the adult attachment sites are defined as early as the third larval instar. Both anterior and posterior sites of attachment of the IFMs are marked by the expression of reporter beta-galactosidase activity in a P-element line B14.0, which is an insertion at the stripe locus. B14.0 (stripe) is seen in distinct domains in the wing and leg imaginal discs which give rise to the thoracic cuticle. The expression is maintained during pupal development. The B14.0 (stripe) expressing epidermal cells contact the developing muscle fibers, leading to the formation of the myotendon junction. We show that the dorsal and ventral attachment sites of one group of IFMs, the DVMs arise from two different imaginal discs (wing and leg, respectively), which may explain the differential effect of mutations such as bendless on these muscles. Attachment sites for the other group of IFMs, the DLMs, on the other hand, arise from one imaginal disc (wing). B14.0 (stripe) expression defines epidermal cells of the adult attachment sites and is likely to function during early events leading to the formation of muscle-epithelial contacts. The PS integrins are detected at later stages, suggesting a role in the stabilization and maturation of the muscle-epidermal contacts into myotendon junctions.
Assuntos
Proteínas de Drosophila , Drosophila melanogaster/crescimento & desenvolvimento , Drosophila melanogaster/genética , Genes de Insetos , Integrinas/fisiologia , Desenvolvimento Muscular , Animais , Proteínas de Ligação a DNA/genética , Drosophila melanogaster/metabolismo , Voo Animal , Imuno-Histoquímica , Cadeias alfa de Integrinas , Integrinas/imunologia , Larva/metabolismo , Metamorfose Biológica , Músculos/metabolismo , Junção Neuromuscular/metabolismo , Fatores de Transcrição/genéticaRESUMO
The bithorax complex (BX-C) of Drosophila, one of two complexes that act as master regulators of the body plan of the fly, has now been entirely sequenced and comprises approximately 315,000 bp, only 1.4% of which codes for protein. Analysis of this sequence reveals significantly overrepresented DNA motifs of unknown, as well as known, functions in the non-protein-coding portion of the sequence. The following types of motifs in that portion are analyzed: (i) concatamers of mono-, di-, and trinucleotides; (ii) tightly clustered hexanucleotides (spaced < or = 5 bases apart); (iii) direct and reverse repeats longer than 20 bp; and (iv) a number of motifs known from biochemical studies to play a role in the regulation of the BX-C. The hexanucleotide AGATAC is remarkably overrepresented and is surmised to play a role in chromosome pairing. The positions of sites of highly overrepresented motifs are plotted for those that occur at more than five sites in the sequence, when < 0.5 case is expected. Expected values are based on a third-order Markov chain, which is the optimal order for representing the BXCALL sequence.
Assuntos
Drosophila/genética , Genes de Insetos , Sequências Reguladoras de Ácido Nucleico , Sequência de Aminoácidos , Animais , Sequência de Bases , DNA/genética , DNA/metabolismo , Proteínas de Ligação a DNA/genética , Dados de Sequência Molecular , Análise de Sequência de DNARESUMO
In Drosophila the Abdominal-B (Abd-B) domain of the bithorax complex (BX-C) spans over 100 kb and is responsible for specifying the identities of adult abdominal segments five (A5) to nine (A9), inclusive, and correspondingly, neuromeres 10-14 of the embryonic central nervous system. The domain consists of a region coding for two proteins, ABD-BI (54 kd) and ABD-BII (36 kd) and cis-regulatory regions extending from infra-abdominal-5 (iab-5) to iab-9, inclusive. We have used a monoclonal anti-ABD-B antibody to infer that mutants in iab-8 eliminate the expression of ABD-BI in neuromeres 10-13, inclusive, and that mutants in iab-9 eliminate expression of ABD-BII in neuromere 14. ABD-B expression is also analyzed in homozygotes for (i) loss-of-function mutants involving the iab-5, iab-6 and iab-7 regions, (ii) gain-of-function mutants Miscadastral pigmentation (Mcp) and Superabdominal (Sab), and (iii) a trans-regulator, Polycomb (Pc). ABD-B expression along the antero-posterior axis is colinear with the chromosomal order of the cis-regulatory regions. The behavior of rearrangement-associated iab-6 and iab-7 mutants suggests that the enhancer-like region, iab-5, and possibly also iab-6, may be shared between the abd-A and Abd-B domains. Such sharing is proposed as a factor that tends to keep gene complexes intact during evolution.
Assuntos
Abdome , Proteínas de Drosophila , Drosophila/genética , Cabeça , Proteínas de Homeodomínio , Hormônios de Inseto/genética , Sequências Reguladoras de Ácido Nucleico , Animais , Cromossomos/metabolismo , Drosophila/embriologia , Regulação da Expressão Gênica , Homozigoto , Hormônios de Inseto/biossíntese , Masculino , MutaçãoRESUMO
In Drosophila, stripe (sr) gene function is required for normal muscle development. Some mutations disrupt embryonic muscle development and are lethal. Other mutations cause total loss of only a single muscle in the adult. Molecular analysis shows that sr encodes a predicted protein containing a zinc finger motif. This motif is homologous to the DNA binding domains encoded by members of the early growth response (egr) gene family. In mammals, expression of egr genes is induced by intercellular signals, and there is evidence for their role in many developmental events. The identification of sr as an egr gene and its pattern of expression suggest that it functions in muscle development via intercellular communication.
Assuntos
Proteínas de Ligação a DNA/biossíntese , Proteínas de Drosophila , Drosophila melanogaster/fisiologia , Genes de Insetos , Músculos/fisiologia , Fatores de Transcrição/biossíntese , Sequência de Aminoácidos , Animais , Sequência de Bases , Elementos de DNA Transponíveis , Proteínas de Ligação a DNA/química , Proteínas de Ligação a DNA/genética , Drosophila melanogaster/embriologia , Drosophila melanogaster/genética , Embrião não Mamífero , Expressão Gênica , Biblioteca Genômica , Hibridização In Situ , Mamíferos , Dados de Sequência Molecular , Músculos/embriologia , Mutagênese Insercional , Mutação , Proteínas Recombinantes/biossíntese , Glândulas Salivares/citologia , Glândulas Salivares/metabolismo , Homologia de Sequência de Aminoácidos , Fatores de Transcrição/química , Fatores de Transcrição/genética , Transcrição Gênica , beta-Galactosidase/biossínteseRESUMO
The bithorax complex (BX-C) of Drosophila, one of two complexes that act as master regulators of the body plan of the fly, is included within a sequence of 338,234 bp (SEQ89E). This paper presents the strategy used in sequencing SEQ89E and an analysis of its open reading frames. The BX-C sequence (BXCALL) contains 314,895 bp obtained by deletion of putative genes that are located at each end of SEQ89E and appear to be functionally unrelated to the BX-C. Only 1.4% of BXCALL codes for the three homeodomain-containing proteins of the complex. Principal findings include a putative ABD-A protein (ABD-AII) larger than a previously known ABD-A protein and a putative glucose transporter-like gene (1521 bp) located at or near the bithoraxoid (bxd), infra-abdominal-2 (iab-2) boundary on the opposite strand relative to that of the homeobox-containing genes.