[Construction and identification of recombinant adenovirus of muramidase-released protein gene fragment from Streptococcus suis type 2].
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi
; 29(4): 350-3, 2013 Apr.
Article
em Zh
| MEDLINE
| ID: mdl-23643163
ABSTRACT
OBJECTIVE:
To construct and identify the recombinant adenovirus of muramidase-released protein (MRP) gene fragment from Streptococcus suis type 2 (SS2).METHODS:
The specific primers were designed based on the sequence of MRP gene fragment. The MRP gene fragment (467-1351 bp) was amplified by PCR method with genomic DNA of SS2 as a template. PCR products were cloned in pMD18-T vector. Then MRP gene fragment was linked into the adenovirus shuttle plasmid (pShuttle-CMV) to construct recombinant shuttle plasmid (pShuttle-CMV-MRP). After PmeI digestion, it was transformed into BJ5183-AD-1 competent cells containing adenoviral backbone plasmid pAdEasy-1 to construct homogeneous recombinant adenovirus plasmid (pAdeno-CMV-MRP). Then the recombinant adenovirus plasmid was linearized by PmeI and then transfected into AD-293 cells for viral packaging. Finally, the virus liquid was tested by PCR and Western blotting.RESULTS:
Cytopathic effect (CPE) was observed at 8 d after transfection of linear pAdeno-CMV-MRP in AD-293 cells. MRP gene fragment and protein expression were also detected in the virus liquid.CONCLUSION:
The recombinant adenovirus of MRP gene fragment (rAdeno-MRP) from SS2 was constructed successfully.
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Base de dados:
MEDLINE
Assunto principal:
Proteínas de Bactérias
/
Adenoviridae
/
Streptococcus suis
/
Antígenos de Bactérias
Tipo de estudo:
Diagnostic_studies
Limite:
Humans
Idioma:
Zh
Ano de publicação:
2013
Tipo de documento:
Article