title: [Construction and identification of recombinant adenovirus of muramidase-released protein gene fragment from Streptococcus suis type 2].
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.
  
  
  CONCLUSIONS
  The recombinant adenovirus of MRP gene fragment (rAdeno-MRP) from SS2 was constructed successfully.
authors: Department of Pathogenic Biology, College of Basic Medicine, Jiujiang University, China. comwangtaocom@sohu.com
authors: Tao
authors: T
authors: Wang
authors: Zihao
authors: Z
authors: Tang
authors: Xiaoou
authors: X
authors: Zhou
authors: Hui
authors: H
authors: Yu
authors: Yingtang
authors: Y
authors: Zhou
mesh_terms: D000256:Adenoviridae / Q000235:genetics* / Q000378:metabolism*; D000942:Antigens, Bacterial / Q000235:genetics* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D002460:Cell Line; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017011:Streptococcus suis / Q000235:genetics* / Q000378:metabolism*; D014162:Transfection / Q000379:methods
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; C421489:MRP protein, Streptococcus suis; D011993:Recombinant Fusion Proteins