Establishment of HEK293 cell line expressing sense and antisense Alu-Sx stably and efficiently
- VernacularTitle:稳定高效表达正反义Alu-Sx细胞系的建立
- Author:
Liang PENG
;
Gang WU
;
Jizhi JIANG
;
Fengshuo JIN
;
Rui DING
- Publication Type:Journal Article
- Keywords:
Alu subfamily Sx;
eukaryotic expressing vector;
HEK293 cell line;
stable transfection
- From:Journal of Third Military Medical University
1988;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a HEK293 cell line stably and highly expressing sense and antisense Alu-Sx.Methods According to Alu subfamily Sx sequence,a pair of primers containing the sites for given restrictive endonuclease at both ends were designed and synthesized.PCR of the total DNA extracted from HEK293 cell line was performed,the products of which were cloned into a highly efficient eukaryotic expression vector pcDNA3.1/myc-His A.The recombinants were sequenced and identified by restrictive endonuclease digestion and then transfected into the HEK293 cell line by lipofectamine2000.The stable transfectants were screened by G418.Cell subclones were isolated by gradient dilution.The highly expressing clones were identified by Northern blotting.Results Eukaryotic expressing vectors stably and efficiently expressing sense and antisense Alu-Sx were constructed and cell subclones stably and efficiently expressing sense and antisense Alu-Sx were established.Conclusion Cell subclones stably and efficiently expressing sense and antisense Alu-Sx can used for our further study.
