Construction and identification of Kir2ds4 RNAi lentiviral vector / 中国实验血液学杂志
Journal of Experimental Hematology
; (6): 663-666, 2008.
Article
in Zh
| WPRIM
| ID: wpr-267915
Responsible library:
WPRO
ABSTRACT
This study was aimed to construct a lentiviral vector of RNA interfered (RNAi)-kir2ds4 gene. In accordance with study-confirmed effective sequence of siRNA targeting kir2ds4 gene, the complementary DNA containing both sense and antisense oligonuctide of the targeting sequence was designed, synthesized and inserted into pSicoR-GFP vector containing U6 promoter and GFP sequence. The resulting lentiviral vector containing kir2ds4 shRNA was named as LV-sh kir2ds4, and confirmed by PCR and sequencing. 293T cells were co-transfected with lentiviral vector LV-sh kir2ds4 and packaging system. All virus stocks were produced by Lipofectamine 2000-mediated transfection. The titer of virus was tested according to the expression level of GFP. As a result, PCR and DNA sequencing demonstrated that the lentivirus RNAi vector of kir2ds4 was constructed successfully. The titer of virus tested by expression level of GFP was 6 x 10(8) TU/ml. It is concluded that the lentivirus RNAi vector of kir2ds4 has been successfully constructed.
Full text:
1
Index:
WPRIM
Main subject:
Molecular Sequence Data
/
Base Sequence
/
Lentivirus
/
RNA, Small Interfering
/
RNA Interference
/
Green Fluorescent Proteins
/
Receptors, KIR
/
Genetic Vectors
/
Genetics
/
Metabolism
Limits:
Humans
Language:
Zh
Journal:
Journal of Experimental Hematology
Year:
2008
Type:
Article