title: [Retroviral vector-mediated HBsAg expression and its stability].
abstract: OBJECTIVE
  To explore use of retroviral vector in gene therapy of hepatitis B.
  
  
  METHODS
  The recombinant vector Plxsn-HBs was constructed by inserting HBV S gene into pLXSN. The pseudovirus, which was produced from PA317 after transferring with pLXSN-HBs by electroporation, were frozen at different temperature. The activities of the pseudovirus to infect eukaryotic cells and express antigen were determined by comparing the numbers of G418-resistant clones and assaying HBsAg in the supernatant of the cells with ELISA after infection HepG2, NIH3T3 and 293 cells.
  
  
  RESULTS
  It was hard to find changes in HBsAg amount at different intervals and different temperatures. G418 resistant clones, however, were variable. When frozen at -20 degrees C, the numbers of clones were half less than that of the beginning after 6 months, few clones were formed after 12 months, and no clone was found after 24 months. When frozen at -40 degrees C, the numbers of clones were 121, 332 and 89 42, 137 and 43 for HepG2, NIH3T3 and 293 cell lines at 12 and 24 months, respectively. When frozen at -70 degrees C, the numbers of clones were 159 463 and 112 for HepG2, NIH3T3 and 293 cell lines at 24 months, respectively. There was no statistical difference compared to that of zero months.
  
  
  CONCLUSIONS
  The activity of the peseudovirus to infect eukaryotic cells and expressed antigen was not changed after 2 years frozen at -70 degrees C.
authors.affiliation: Infectious Disease Department, The Second Affiliated Hospital, Chongqing Medical University, Chongqing 400010, China.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Zhou
authors.forename: Haihong
authors.initials: H
authors.lastname: Zhang
authors.forename: Jianxi
authors.initials: J
authors.lastname: Lu
authors.forename: Jilu
authors.initials: J
authors.lastname: Yao
authors.forename: Lainxian
authors.initials: L
authors.lastname: Deng
mesh_terms: D002460:Cell Line; D005822:Genetic Vectors; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis* / Q000235:genetics; D012190:Retroviridae / Q000235:genetics*; D013696:Temperature; D014162:Transfection
chemical_list: D006514:Hepatitis B Surface Antigens