title: [The non-replicating recombinant vaccinia virus expressing six genes of HIV-1 can be passaged stably in CEF].
abstract: To investigate the genetic stability (including the vector of vaccinia virus and six foreign genes: gp160, gag, pol, rev, tat and nef) of the HIV-1 non-replicating recombinant vaccinia virus (rNTV-C). rNTV-C was serially passaged to passage 25 (P25) in primary chicken embryo fibroblast (CEF). P9, P12, P15 and P25 were selected to study the genetic stability in four aspects, including the genetic stability of viral vector, the genetic stability of six foreign genes, the expressing stability of foreign genes and the genetic loss of foreign genes. The results showed that the viral vector was non-replicated vaccinia virus of Tiantan strain and was passaged stably; foreign gene sequences matched with designed sequences, the insert sites were right, and the nucleotide mutation rate was less than one over ten thousands within different passages of rNTV-C; the target proteins could be expressed effectively, and the expression level was stable within different passages of rNTV-C; the genetic loss of gag and nef was less than 5% within different passages of rNTV-C. The above results provided important data for the vaccine production.
authors.affiliation: Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China. qxryyw@163.com
authors.forename: Xiang-rong
authors.initials: XR
authors.lastname: Qi
authors.forename: Xiang-min
authors.initials: XM
authors.lastname: Zhang
authors.forename: Yao
authors.initials: Y
authors.lastname: Deng
authors.forename: Ying-ying
authors.initials: YY
authors.lastname: Gao
authors.forename: Rou-jian
authors.initials: RJ
authors.lastname: Lu
authors.forename: Xin
authors.initials: X
authors.lastname: Meng
authors.forename: Wen-jie
authors.initials: WJ
authors.lastname: Tan
authors.forename: Li
authors.initials: L
authors.lastname: Ruan
mesh_terms: D000818:Animals; D004274:DNA, Recombinant / Q000235:genetics*; D005347:Fibroblasts / Q000378:metabolism* / Q000821:virology*; D015870:Gene Expression; D005814:Genes, Viral / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D015497:HIV-1 / Q000235:genetics*; D017422:Sequence Analysis, DNA; D014616:Vaccinia virus / Q000235:genetics*
chemical_list: D004274:DNA, Recombinant