No description available.

The strategy of producing expressed sequence tags (ESTs) is greatly used for gene mining and analysis of gene expression. An EST is a partial and single-pass sequence generated from a complementary DNA (cDNA) library by random selection, and is typically about 400 to 600 bases. Here, we constructed and analyzed a brain EST library from 10-day-old (10-day posthatching) tilapia, Oreochromis mossambicus, to assist in investigating the relation between brain neural development and neuroendocrine and brain sexual differentiation during the critical period of sexual dimorphism by understanding brain gene expression, and, furthermore, to find genes that are important in brain but are unknown nowadays. A total 1,124 ESTs were in the 10-day-old tilapia brain EST library, and 1,092 ones were readable, and after discarding one that is too short (<200bp) or is totally vector sequence from the readable ESTs, 1,084 ESTs were assembled and then analyzed. 108 contigs from 261 ESTs, 20 kinds of similar sequences from 40 ESTs, and 783 singletons from the 1,084 ESTs were found, and after aligning in turn with the non-redundant (nr) protein database, nr nucleotide database and EST database dbEST supported by the National Center of Biotechnology Information (NCBI) using Basic Local Alignment Search Tool (BLAST) programs BLASTx (translated nucleotide-protein alignment) and BLASTn (nucleotide-nucleotide alignment) respectively, the results are as follows: 57 and 16 contigs including 146 and 37 ESTs matched with sequences in the nr protein and nr nucleotide databases respectively, and the rest 35 contigs including 78 ESTs were novel sequences; nine, one and two kinds of the similar sequences including 18, two and four ESTs matched with sequences in the nr protein, nr nucleotide and dbEST databases respectively, and the rest eight kinds of the similar sequences including 16 ESTs were novel sequences; 309, 52 and 47 singletons matched with sequences in the nr protein, nr nucleotide and dbEST databases respectively, and the rest 375 singletons were novel sequences. As a whole, the most functions of proteins that the contigs, similar kinds of sequences, and singletons from the 10-day-old tilapia brain EST library matched with were binding and transport activity. Recently, many researchers provide reasonable explanations for evolution, relation between genomic polymorphism and drug effects, and isoforms presentation of known proteins by collecting ESTs from open EST databases or from EST libraries they constructed, and we believe that this 10-day-old tilapia brain EST library can promote our ability to resolve questions about the topic we are researching in.

O carrapato Boophilus microplus, um ectoparasita hematófago, está presente em áreas tropicais e subtropicais no mundo todo. Atualmente, seu controle é baseado no uso de pesticidas. Para auxiliar no desenvolvimento de novas vacinas para este parasita, é importante compreender melhor sua fisiologia. A coronina, uma proteína de citoesqueleto, pertence à Família das Proteínas G. Está localizada no córtex celular e apresenta várias funções, desde locomoção celular, fagocitose, macropinocitose a ligação de microtúbulos, orientação da polaridade de embriões, citocinese, polimerização dos filamentos de actina e regulação da organização do citoesqueleto. No presente trabalho, o cDNA de uma proteína semelhante a coronina foi isolado de uma biblioteca de cDNA de glândulas salivares de partenóginas por triagem imunológica. A análise de seqüência mostrou que o clone SG5 possui identidade entre 15 e 17% com seqüências de proteínas semelhantes a coronina de outras espécies, como as de humanos e de camundongo, e possui um tamanho de 1.938 pb, com uma suposta ORF de 960 nucleotídeos. Na análise da expressão do gene, verificouse que o mRNA deste gene está presente em ovos e larvas de 15 dias e glândulas salivares, intestino e ovário de teleógina e partenógina; em corpo gorduroso sua presença não foi detectada. Um estudo mais detalhado do papel da proteína semelhante a coronina nas glândulas salivares de B. microplus pode auxiliar em uma melhor compreensão da fisiologia deste parasita.

O carrapato Boophilus microplus, um ectoparasita hematófago, está presente em áreas tropicais e subtropicais no mundo todo. Atualmente, seu controle é baseado no uso de pesticidas. Para auxiliar no desenvolvimento de novas vacinas para este parasita, é importante compreender melhor sua fisiologia. A coronina, uma proteína de citoesqueleto, pertence à Família das Proteínas G. Está localizada no córtex celular e apresenta várias funções, desde locomoção celular, fagocitose, macropinocitose a ligação de microtúbulos, orientação da polaridade de embriões, citocinese, polimerização dos filamentos de actina e regulação da organização do citoesqueleto. No presente trabalho, o cDNA de uma proteína semelhante a coronina foi isolado de uma biblioteca de cDNA de glândulas salivares de partenóginas por triagem imunológica. A análise de seqüência mostrou que o clone SG5 possui identidade entre 15 e 17% com seqüências de proteínas semelhantes a coronina de outras espécies, como as de humanos e de camundongo, e possui um tamanho de 1.938 pb, com uma suposta ORF de 960 nucleotídeos. Na análise da expressão do gene, verificouse que o mRNA deste gene está presente em ovos e larvas de 15 dias e glândulas salivares, intestino e ovário de teleógina e partenógina; em corpo gorduroso sua presença não foi detectada. Um estudo mais detalhado do papel da proteína semelhante a coronina nas glândulas salivares de B. microplus pode auxiliar em uma melhor compreensão da fisiologia deste parasita.

O carrapato Boophilus microplus, um ectoparasita hematófago, está presente em áreas tropicais e subtropicais no mundo todo. Atualmente, seu controle é baseado no uso de pesticidas. Para auxiliar no desenvolvimento de novas vacinas para este parasita, é importante compreender melhor sua fisiologia. A coronina, uma proteína de citoesqueleto, pertence à Família das Proteínas G. Está localizada no córtex celular e apresenta várias funções, desde locomoção celular, fagocitose, macropinocitose a ligação de microtúbulos, orientação da polaridade de embriões, citocinese, polimerização dos filamentos de actina e regulação da organização do citoesqueleto. No presente trabalho, o cDNA de uma proteína semelhante a coronina foi isolado de uma biblioteca de cDNA de glândulas salivares de partenóginas por triagem imunológica. A análise de seqüência mostrou que o clone SG5 possui identidade entre 15 e 17% com seqüências de proteínas semelhantes a coronina de outras espécies, como as de humanos e de camundongo, e possui um tamanho de 1.938 pb, com uma suposta ORF de 960 nucleotídeos. Na análise da expressão do gene, verificouse que o mRNA deste gene está presente em ovos e larvas de 15 dias e glândulas salivares, intestino e ovário de teleógina e partenógina; em corpo gorduroso sua presença não foi detectada. Um estudo mais detalhado do papel da proteína semelhante a coronina nas glândulas salivares de B. microplus pode auxiliar em uma melhor compreensão da fisiologia deste parasita.

A 5' cDNA clone coding for human C4b-binding protein (C4bp) was isolated, characterised and sequenced to complete the cDNA sequence coding for amino-acid residues 1-32 thus confirming the protein sequence data of Chung et al. (1985c). The sequence extended to allow derivation of the putative leader sequence which was 32 residues in length and showed a high degree of hydrophobicity typical of other documented leader sequences. This clone together with others isolated by Chung et al. (1985a) were used to prepare cDNA probes which were subsequently used to hybridise genomic clones. The data from the blots suggested that the C4bp gene was up to 30 kb in size and thus consisted of large proportions of intron sequence (mRNA ≃ 2.5 kb). A commonly occurring restriction fragment length polymorphism was detected using the BglII restriction enzyme. Analysis of the genomic clones has shown that, with one possible exception, each internal protein repeat is encoded by a discrete exon. Use of the cDNA probes to hybridise RNA transfer blots, on which the RNA originated from various tissues and cell-lines, suggested that the liver is the major source of C4bp mRNA. Cross hybridisation was detected between the human C4bp cDNA probes and genomic DNA isolated from various species on transfer blots suggesting that genomic sequence homologous to that coding for C4bp has been conserved during evolution. Results from pulse field gel electrophoresis blot analysis suggests that although C4bp and the alternative pathway regulatory protein, Factor H, are genetically linked the physical distance separating them may be as great as 3 Mbp.

No description available.

No description available.

Cultivated chickpea (Cicer arietinum) has a narrow genetic base making it difficult for breeders to produce new elite cultivars with durable resistance to major biotic and abiotic stresses. As an alternative to genome mapping, microarrays have recently been applied in crop species to identify and assess the function of putative genes thought to be involved in plant abiotic stress and defence responses. In the present study, a cDNA microarray approach was taken in order to determine if the transcription of genes, from a set of previously identified putative stress-responsive genes from chickpea and its close relative Lathyrus sativus, were altered in chickpea by the three abiotic stresses; drought, cold and high-salinity. For this, chickpea genotypes known to be tolerant and susceptible to each abiotic stress were challenged and gene expression in the leaf, root and/or flower tissues was studied. The transcripts that were differentially expressed among stressed an d unstressed plants in response to the particular stress were analysed in the context of tolerant/susceptible genotypes. The transcriptional change of more than two fold was observed for 109, 210 and 386 genes after drought, cold and high-salinity treatments, respectively. Among these, two, 15 and 30 genes were consensually differentially expressed (DE) between tolerant and susceptible genotypes studied for drought, cold and high-salinity, respectively. The genes that were DE in tolerant and susceptible genotypes under abiotic stresses code for various functional and regulatory proteins. Significant differences in stress responses were observed within and between tolerant and susceptible genotypes highlighting the multiple gene control and complexity of abiotic stress response mechanism in chickpea. The annotation of these genes suggests that they may have a role in abiotic stress response and are potential candidates for tolerance/susceptibility.

An infectious cDNA clone of Vero cell adapted Beaudette strain of IBV was constructed using in vitro assembly of cDNA fragments. The entire genome of IBV was RT-PCR amplified into seven fragments, with each piece overlapping about 10 nucleotides. The fragments were ligated and transcribed to synthesize RNA, which was transfected into BHK-21 cells. These cells were then overlaid onto IBV susceptible Vero cells. After five days transfection, the virus was successfully rescued from the transfected cells. The cDNA clone from our laboratory strain has a five nucleotide insertion not present in the originally sequenced virus, resulting in total genome size of 27,613 nucleotides. The infectious cDNA clone was further manipulated to demonstrate its potential as a gene transfer vector, by replacing the ORF5a open reading frame with enhanced green fluorescent protein. The recombinant infectious cDNA clone was also successfully rescued after three days transfection of BHK-21 cells followed by co-culturing with Vero cells. This study showed that the 5a protein, whose function is not known, is not necessary for in vitro IBV replication. This study also showed that the 5a ORF is a good candidate for an insertion site of recombinant genes for the development of IBV infectious cDNA clone as a gene transfer vector.