| Registro completo |
| Provedor de dados: |
Genet. Mol. Biol.
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| País: |
Brazil
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| Título: |
Molecular cloning and expression of key gene encoding hypothetical DNA polymerase from B. mori parvo-like virus
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| Autores: |
Zhang,Junhong
Li,Guohui
Chen,Huiqing
Li,Xiaogang
Lv,Meng
Chen,Keping
Yao,Qin
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| Data: |
2010-01-01
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| Ano: |
2010
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| Palavras-chave: |
Bombyx mori parvo-like virus
DNA polymerase
Recombinant protein
Sf-9
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| Resumo: |
BmPLV-Z is the abbreviation for Bombyx mori parvo-like virus (China isolate). This is a novel virus with two single-stranded linear DNA molecules, viz., VD1 (6543 bp) and VD2 (6022 bp), which are encapsidated respectively into separate virions. Analysis of the deduced amino acid sequence of VD1-ORF4 indicated the existence of a putative DNA-polymerase with exonuclease activity, possibly involved in the replication of BmPLV-Z. In the present study, a recombinant baculovirus was constructed to express the full length of the protein encoded by the VD1-ORF4 gene (3318 bp). In addition, a 2163-bp fragment amplified from the very same gene was cloned into prokaryotic expression vector pET-30a and expressed in E.coli Rosetta 2 (DE3) pLysS. The expressed fusion protein was employed to immunize New Zealand white rabbits for the production of an antiserum, afterwards used for examining the expression of the protein encoded by VD1-ORF4 gene in Sf-9 cells infected with recombinant baculovirus. Western blot analysis of extracts from thus cells infected revealed a specific band of about 120 kDa, thereby indicating that the full length protein encoded by the VD1-ORF4 gene had been successfully and stably expressed in Sf-9 cells.
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| Tipo: |
Info:eu-repo/semantics/article
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| Idioma: |
Inglês
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| Identificador: |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572010000400021
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| Editor: |
Sociedade Brasileira de Genética
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| Relação: |
10.1590/S1415-47572010005000083
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| Formato: |
text/html
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| Fonte: |
Genetics and Molecular Biology v.33 n.4 2010
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| Direitos: |
info:eu-repo/semantics/openAccess
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