RESUMO
The complete nucleotide sequence (155 844 bp) of tobacco (Nicotiana tabacum var. Bright Yellow 4) chloroplast DNA has been determined. It contains two copies of an identical 25 339 bp inverted repeat, which are separated by a 86 684 bp and a 18 482 bp single-copy region. The genes for 4 different rRNAs, 30 different tRNAs, 39 different proteins and 11 other predicted protein coding genes have been located. Among them, 15 genes contain introns. Blot hybridization revealed that all rRNA and tRNA genes and 27 protein genes so far analysed are transcribed in the chloroplast and that primary transcripts of the split genes hitherto examined are spliced. Five sequences coding for proteins homologous to components of the respiratory-chain NADH dehydrogenase from human mitochondria have been found. The 30 tRNAs predicted from their genes are sufficient to read all codons if the ;two out of three' and ;U:N wobble' mechanisms operate in the chloroplast. Two sequences which autonomously replicate in yeast have also been mapped. The sequence and expression analyses indicate both prokaryotic and eukaryotic features of the chloroplast genes.
RESUMO
Transpositions of Alu sequences, representing the most abundant primate short interspersed elements (SINE), were evaluated as molecular cladistic markers to analyze the phylogenetic affiliations among the primate infraorders. Altogether 118 human loci, containing intronic Alu elements, were PCR analyzed for the presence of Alu sequences at orthologous sites in each of two strepsirhine, New World and Old World monkey species, Tarsius bancanus, and a nonprimate outgroup. Fourteen size-polymorphic amplification patterns exhibited longer fragments for the anthropoids (New World and Old World monkeys) and T. bancanus whereas shorter fragments were detected for the strepsirhines and the outgroup. From these, subsequent sequence analyses revealed three Alu transpositions, which can be regarded as shared derived molecular characters linking tarsiers and anthropoid primates. Concerning the other loci, scenarios are represented in which different SINE transpositions occurred independently in the same intron on the lineages leading both to the common ancestor of anthropoids and to T. bancanus, albeit at different nucleotide positions. Our results demonstrate the efficiency and possible pitfalls of SINE transpositions used as molecular cladistic markers in tracing back a divergence point in primate evolution over 40 million years old. The three Alu insertions characterized underpin the monophyly of haplorhine primates (Anthropoidea and Tarsioidea) from a novel perspective.
Assuntos
Haplorrinos/fisiologia , Elementos Nucleotídeos Curtos e Dispersos , Tarsiidae/genética , Tarsiidae/fisiologia , Animais , Sequência de Bases , Bases de Dados Factuais , Marcadores Genéticos , Haplorrinos/classificação , Haplorrinos/genética , Humanos , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase , Polimorfismo Genético , Análise de Sequência de DNA , Homologia de Sequência do Ácido Nucleico , Software , Tarsiidae/classificaçãoRESUMO
DNA sequencing has revealed a long open reading frame (ORF) in the large single-copy region of tobacco chloroplast DNA. This ORF consists of 1070 codons and its deduced amino acid sequence shows about 39% homology to that of the beta-subunit of E. coli RNA polymerase. This finding raises a possibility that some of the chloroplast RNA polymerase subunits are coded for by the chloroplast genome.
Assuntos
Cloroplastos/metabolismo , RNA Polimerases Dirigidas por DNA/genética , Escherichia coli/genética , Peptídeos/genética , Proteínas de Plantas/genética , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , DNA/isolamento & purificação , Escherichia coli/enzimologia , Substâncias Macromoleculares , Plantas/metabolismo , Plantas Tóxicas , Homologia de Sequência do Ácido Nucleico , Nicotiana/metabolismoRESUMO
A non-uniform equivalent cable model of membrane voltage changes in branching neuronal trees with active ion channels has been developed. A general branching condition is formulated, extending Rall's 3/2 power rule for passive dendritic trees so that non-uniform cable segments can be treated. The theoretical results support the use of the dendritic profile model of Clements and Redman. The theory is then applied to dendrites of different morphological type yielding qualitative different response behaviour.
Assuntos
Redes Neurais de Computação , Neurônios/metabolismo , Simulação por Computador , Cibernética , Dendritos/metabolismo , Dendritos/ultraestrutura , Canais Iônicos/metabolismo , Potenciais da Membrana , Modelos Neurológicos , Neurônios/ultraestruturaRESUMO
We have isolated and characterized the genomic clone lambda CHN50 corresponding to tobacco basic endochitinase (E.C.3.2.1.14). DNA sequence and blotting analysis reveal that the coding sequence of the gene present on lambda CHN50 is identical to that of the cDNA clone pCHN50 and, moreover, the CHN50 gene has its origin in the progenitor of tobacco, Nicotiana sylvestris. Tobacco basic chitinases are encoded by a small gene family that consists of at least two members, the CHN50 gene and a closely related CHN17 gene which was characterized previously. By northern blot analysis, it is shown that the CHN50 gene is highly expressed in suspension-cultured tobacco cells and the mRNA accumulates at late logarithmic growth phase. To identify cis-DNA elements involved in the expression of the CHN50 gene in suspension-cultured cells, the chimeric gene consisting of 1.1 kb CHN50 5' upstream region fused to the coding sequence of beta-glucuronidase (GUS) was introduced by electroporation into protoplasts isolated from suspension-cultured tobacco cells. Transient GUS activity was found to be dependent on the growth phase of the cultured cells, from which protoplasts had been prepared. Functional analysis of 5' deletions suggests that the distal region between -788 and -345 contains sequences that potentiate the high-level expression in tobacco protoplasts and the region (-68 to -47) proximal to the TATA box functions as a putative silencer.
Assuntos
Quitinases/genética , Nicotiana/genética , Plantas Tóxicas , Sequência de Aminoácidos , Sequência de Bases , Células Cultivadas , Quitinases/biossíntese , Clonagem Molecular , Regulação Enzimológica da Expressão Gênica , Glucuronidase/genética , Dados de Sequência Molecular , Regiões Promotoras Genéticas , Nicotiana/enzimologiaRESUMO
The complete mitochondrial genome of Tupaia belangeri, a representative of the eutherian order Scandentia, was determined and compared with full-length mitochondrial sequences of other eutherian orders described to date. The complete mitochondrial genome is 16, 754 nt in length, with no obvious deviation from the general organization of the mammalian mitochondrial genome. Thus, features such as start codon usage, incomplete stop codons, and overlapping coding regions, as well as the presence of tandem repeats in the control region, are within the range of mammalian mitochondrial (mt) DNA variation. To address the question of a possible close phylogenetic relationship between primates and Tupaia, the evolutionary affinities among primates, Tupaia and bats as representatives of the Archonta superorder, ferungulates, guinea pigs, armadillos, rats, mice, and hedgehogs were examined on the basis of the complete mitochondrial DNA sequences. The opossum sequence was used as an outgroup. The trees, estimated from 12 concatenated genes encoded on the mitochondrial H-strand, add further molecular evidence against an Archonta monophyly. With the new data described in this paper, most of both the mitochondrial and the nuclear data point away from Scandentia as the closest extant relatives to primates. Instead, the complete mitochondrial data support a clustering of Scandentia with Lagomorpha connecting to the branch leading to ferungulates. This closer phylogenetic relationship of Tupaia to rabbits than to primates first received support from several analyses of nuclear and partial mitochondrial DNA data sets. Given that short sequences are of limited use in determining deep mammalian relationships, the partial mitochondrial data available to date support this hypothesis only tentatively. Our complete mitochondrial genome data therefore add considerably more evidence in support of this hypothesis.
Assuntos
DNA Mitocondrial/genética , Filogenia , Escandêntias/genética , Tupaia/genética , Animais , Sequência de Bases , DNA Mitocondrial/química , Mitocôndrias/genética , Mitocôndrias/metabolismo , Dados de Sequência Molecular , Proteínas/genética , RNA Ribossômico/genética , RNA Ribossômico 16S/genética , RNA de Transferência/genética , Escandêntias/classificação , Análise de Sequência de DNARESUMO
The location and nucleotide sequences of tobacco chloroplast genes for tRNAGlu(UUC), tRNATyr(GUA) and tRNAAsp(GUC) have been determined. These genes lie midway between the genes for alpha and beta/epsilon subunits of H+-ATPase on the large single-copy region of the chloroplast DNA. The gene organization is tRNAGlu - 59bp spacer - tRNATyr - 108bp spacer - tRNAAsp on the same DNA strand. Northern blot hybridization studies revealed that these three tRNA genes are cotranscribed. The transcription initiation site was localized at 24 bp upstream from the tRNAGlu coding region and its termination site at 90 bp downstream from the tRNAAsp coding region by S1 mapping. The tricistronic tRNA precursor is thus calculated to be 512 bases long. Its processing was also studied by S1 mapping.
Assuntos
Cloroplastos/fisiologia , Nicotiana/genética , Plantas Tóxicas , RNA de Transferência/genética , Sequência de Bases , Códon , Regulação da Expressão Gênica , Genes , Ligação Genética , Conformação de Ácido Nucleico , Precursores de Ácido Nucleico/genética , Óperon , Processamento Pós-Transcricional do RNA , Transcrição GênicaRESUMO
The location and nucleotide sequences of tobacco chloroplast genes for tRNA(Ile) (CAU), tRNA(Leu) (CAA), tRNA(Cys) (GCA), tRNA(Ser) (UGA) and tRNA(Thr) (GGU) (trnI-CAU, trnL-CAA, trnC-GCA, trnS-UGA and trnT-GGU, respectively) have been determined. The trnI and trnL are located in the inverted repeat region. The trnC, trnS and trnT are present in the large single copy region. These five tRNA genes together with the 25 different tRNA genes previously published have been compiled and compared. These 30 tRNA genes corresponding to 20 amino acids are most likely to be all of the tRNA genes encoded in tobacco chloroplast genome.
RESUMO
The nucleotide sequences of tobacco chloroplast genes for tRNAPro(UGG), tRNATrp, tRNAfMet and tRNAGly(GCC) have been determined. None of these genes contains an intron. One unusual feature is that the tRNAGly contains only two base-pairs (A-U, G-U) in the D stem. These four tRNA genes were located in the known physical map of tobacco chloroplast DNA. Hybridization analysis to chloroplast tRNA revealed that all four tRNA genes are transcribed in vivo.
Assuntos
Cloroplastos/análise , Genes , Plantas/genética , RNA de Transferência de Metionina , RNA de Transferência/genética , Composição de Bases , Sequência de Bases , Clonagem Molecular , Enzimas de Restrição do DNA , Conformação de Ácido Nucleico , Plantas Tóxicas , Aminoacil-RNA de Transferência/genética , Nicotiana/genética , Transcrição GênicaRESUMO
Transcription of the psaA operon in tobacco chloroplasts has been studied. This operon contains in linear sequence the genes encoding the P700 chlorophyll a A1 and A2 apoproteins (psaA and psaB) and the gene encoding the ribosomal CS14 protein (rps14). Northern blot hybridization revealed that a 5.2 kb transcript hybridizes to psaA, psaB, and rps14, but not to the fMet-tRNA (trnfM) gene which follows. Primer extension and in vitro capping assays indicated that the transcriptional initiation site is 194 bp upstream of psaA. The 3' end of the transcript was determined by S1 mapping to be 105 bp downstream of rps14. The transcript is calculated to be 5,207 nucleotides long.