title: Construction of conditionally replicating adenovirus expressing staphylococcal enterotoxin A gene: potential usefulness for anti-tumor therapies.
abstract: OBJECTIVE
  The aim of this study was to construct a conditionally replicating adenovirus pPE3-SEA expressing staphylococcal enterotoxin A (SEA) gene.
  
  
  METHODS
  A full-length SEA gene fragment was cloned into pENTR12 plasmid to obtain a recombinant viral plasmid pENTR12-SEA. The pENTR12-SEA plasmid was co-transfected into HEK293 cells along with pPE3-ccdB, which encoded for the virus backbone, to generate recombinant adenovirus pPE3-SEA vector. Amplified pPE3-SEA vectors were purified, and viral titer was determined using the 50% tissue culture infective dose method.
  
  
  RESULTS
  The PCR, restriction enzyme digestion, and sequence analyses proved successful construction of replicating oncolytic adenovirus pENTR12-SEA and recombinant SEA expressing oncolytic adenovirus pPE3-SEA. The viral titer was 2.5 × 1010 pfu/ml.
  
  
  CONCLUSIONS
  We successfully constructed conditionally replicating adenovirus pPE3-SEA which can be utilized for experimental studies of tumor-targeted therapies.
authors.affiliation: Department of Urology, The Afﬁliated School of Clinical Medicine of Xuzhou Medical College, Xuzhou Central Hospital, Xuzhou, China. 1312552260@qq.com.
authors.forename: P-Y
authors.initials: PY
authors.lastname: Zhang
authors.forename: L
authors.initials: L
authors.lastname: Hao
authors.forename: Z-G
authors.initials: ZG
authors.lastname: Zhang
authors.forename: B-Z
authors.initials: BZ
authors.lastname: Dong
authors.forename: D
authors.initials: D
authors.lastname: Yang
authors.forename: X-L
authors.initials: XL
authors.lastname: Wang
authors.forename: X-J
authors.initials: XJ
authors.lastname: Xuan
authors.forename: Z
authors.initials: Z
authors.lastname: Yan
authors.forename: L
authors.initials: L
authors.lastname: Qing
authors.forename: Z-D
authors.initials: ZD
authors.lastname: Shi
authors.forename: D
authors.initials: D
authors.lastname: Liu
authors.forename: C-H
authors.initials: CH
authors.lastname: Han
mesh_terms: D000256:Adenoviridae / Q000235:genetics* / Q000502:physiology; D004768:Enterotoxins / Q000235:genetics*; D015316:Genetic Therapy; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D009369:Neoplasms / Q000628:therapy; D014162:Transfection; D014779:Virus Replication
chemical_list: D004768:Enterotoxins; C026066:enterotoxin A, Staphylococcal