[Retroviral vector-mediated HSVtk gene expression and acquisition of high titer recombinant virus].
OBJECTIVE
To explore the HSVtk gene expression mediated by the retroviral vector and to obtain high titer recombinant retroviral virus.


METHODS
The recombinant vector pRevTRE/HSVtk was constructed by inserting HSVtk gene into pRevTRE. The recombinant retrovirus, which was produced from cloned PA317 cells screened by hygromycin B after "micro-pingpong" technique transferring with pRevTRE/HSVtk plasmids DNA by using modified calcium phosphate precipitation method. HSVtk gene expression was performed on target cells and virus titers were detected in different cultured temper, time and sodium butyrate concentration.


RESULTS
The recombinant retroviral vector pRevTRE/HSVtk was constructed and HSVtk gene expression was detected on target cells after they were infected with the recombinant retrovirus.


CONCLUSIONS
High titer of retroviruses could be obtained in the culture medium of PA317 cell line through "micro-pingpong" technique at 30 hours and 10 mmol/L sodium butyrate concentration followed by frozen ultrafiltration.
Zeng
Zhao-jun
ZJ
Molecular Biology Research Center of Xiangya Medical College, Central South University, Changsha 410078, China.
Hu
Wei-xin
WX
Luo
Sai-qun
SQ
Chen
Qian
Q
D000818:Animals; D001943:Breast Neoplasms / Q000201:enzymology / Q000473:pathology / Q000821:virology; D002460:Cell Line; D045744:Cell Line, Tumor; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D011995:Recombination, Genetic; D012190:Retroviridae / Q000235:genetics*; D018139:Simplexvirus / Q000201:enzymology* / Q000235:genetics; D013937:Thymidine Kinase / Q000096:biosynthesis* / Q000235:genetics; D017186:Titrimetry; D014162:Transfection
D013937:Thymidine Kinase