RESUMO
MicroRNAs (miRNAs) constitute a novel, extensive class of small RNAs (approximately 21 nucleotides), and play important gene-regulation roles during growth and development in various organisms. Here we conducted a homology search to identify homologs of previously validated miRNAs from silkworm genome. We identified 24 potential miRNA genes, and gave each of them a name according to the common criteria. Interestingly, we found that a great number of newly identified miRNAs were conserved in silkworm and Drosophila, and family alignment revealed that miRNA families might possess single nucleotide polymorphisms. miRNA gene clusters and possible functions of complement miRNA pairs are discussed.
Assuntos
Animais , Sequência de Bases , Bombyx , Análise por Conglomerados , Biologia Computacional , Métodos , Drosophila melanogaster , Teste de Complementação Genética , Genoma , MicroRNAs , Metabolismo , Dados de Sequência Molecular , Família Multigênica , Polimorfismo de Nucleotídeo Único , Homologia de Sequência do Ácido Nucleico , Software , TermodinâmicaRESUMO
Segment A of the genome of infectious bursal disease virus(IBDV) encodes structure protein VP2 and VP3 and protease VP4. In this study a polyprotein gene of IBDV was inserted into a Bombyx mori baculovirus transfer vector pAcHLT--C and contransfected into BmN cells with linear genome DNA of virus Bm-BacPAK6. Dot hybridization suggested that the segment A of the virus genome was inserted in the genome of Bm-BacPAK6. The silkworm of fifth instars were infected by the recombinant virus and the immunogenicity of the infected larvae's blood were examined with ELISA, SDS-PAGE and Western blotting. It appears that the recombinant polyprotein has the property of immunoreactivity and the expression in larvae reached the pick 5-day post infection.
Assuntos
Animais , Western Blotting , Bombyx , Genética , Virologia , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Expressão Gênica , Vetores Genéticos , Genética , Vírus da Doença Infecciosa da Bursa , Genética , Poliproteínas , Genética , Metabolismo , Proteínas Virais , Genética , MetabolismoRESUMO
A recombinant transfer vector, pBacIL-11, containing hIL-11 cDNA of 546 nucleotides lacking leader sequence was constructed and co-transfected into BmN cells with linearized BmBacPAK(modified BmNPV) DNA for construction of a recombinant baculovirus carrying the hIL-11 gene. Southern hybridization analysis suggested that the recombinant baculovirus DNA contained hIL-11 cDNA fragment. RNA dot blotting demonstrated that the hIL-11 gene was transcribed. The recombinant baculovirus has a strong infectivity to BmN cell line and to silkworm larvae and pupae. Specific hIL-11 bands were detected from all the samples of cell extract, culture supernatant, haemolymph of larvae and pupae by SDS-PAGE analysis. Biological activity of the expressed product was determined with IL-11 dependent B9-11 cell line and by MTT colorimetric assay, which indicated that biologically active rhIL-11 protein was overexpressed in BmN cell line and in silkworm larvae and pupae.