The Construction and Expression of Human Interleukin-2 Recombinant Retroviral Vector
- VernacularTitle:人白细胞介素2重组逆转录病毒载体的构建与表达
- Author:
Weiping ZHANG
;
Xuetao CAO
;
Jianli WANG
- Publication Type:Journal Article
- Keywords:
Interleukin-2;
retrovirus;
gene cloning;
gene transfer;
gene thrapy
- From:
Chinese Journal of Cancer Biotherapy
1995;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
The human Interleukin-2 cDNA containing full-length of encoding region was cloned by RT-PCR from PBMNC and confirmed by DNA sequencing. The hIL-2 recombinant retroviral expressing vector (pLXSN-hIL2) was constructed by inserting hIL-2 cDNA into the BamHI cloning site of pLXSN retroviral vector. After packaged with CRIP packaging cell line, the hIL-2 retrovirions were produced at the titer of 7.6?105CFU/ml. Then a fibroblast cell clone(NIH3T3-hIL2) secreting 118.2U/ml hIL-2 was obtained by infecting the NIH3T3 fibroblast cells with hIL-2 retrovirions. The integration of hIL-2 provirus into the genome of NIH3T3-ML2 cells was confirmed by PCR analysis for NeoR gene. Our data showed that the hIL-2 retroviral vector was successfully constructed, which enables us to further the investigation of the hIL-2 gene therapy in clinical trial.
