Construction and verification of Ientiviral CRKL gene RNA interfering vector
- VernacularTitle:CRKL基因RNA干扰慢病毒载体的构建与鉴定
- Author:
Shaohua SHEN
;
Aihua LIU
;
Chunhua QI
;
Xin YE
;
Longjun GU
- Publication Type:Journal Article
- Keywords:
RNA interference;
Phosphotyrosine;
Signal transduction
- From:
Journal of International Oncology
2008;35(12):947-950
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a lentiviral vector carrying CRKL gene RNA interfering( RNAi ).Methods The CRKL RNAi was selected and subcloned into the lentiviral vector,pGCL-GFP(including U6 promotor and green fluorescent protein),generating the lentiviral vector LV-shCRKL.The corrected CRKL was confirmed by endoenzyme digestion ,sequencing.Recombinant lentiviruses were produced by 293T cells following the eo-transfection of LV-shCRKL,with the packaging plasmids pHelper1.0 and pHelper2.0.The virus titer was detected by GFP expressions in 293T cells.Results Plasmid LV-shCRKL carried the correct sequence.The recombinant lentiviruse LV-shCRKL could be produced by co-transfection of LV-shCRKL to 293T cells.Conclusion The recombinant lentiviruse vector LV-shCRKL is constructed successful.
