Construction and identification of conditionally replicating adenovirus vector Ad-delE1b55kD-shRNA/Survivin-EGFP / 第三军医大学学报
Journal of Third Military Medical University
;
(24)2003.
Article
in Chinese
| WPRIM
| ID: wpr-561854
ABSTRACT
Objective To construct the conditionally replicating adenovirus vector Ad-delE1b55kD-shRNA/Survivin-EGFP that can transfects into HT-29 effectually and selectively and contains shRNA targeting to human Survivin gene.Methods EGFP geoe and shRNA gene were inserted into pAd-delE1b55kD,then pAd-delE1b55kD-shRNA/Survivin-EGFP was obtained.The plasmid and pBHGE3 were cotransfected into HEK-293 cell to obtain the conditionally replicating adenovirus vector Ad-delE1b55kD-shRNA/Survivin-EGFP.Then DNA of the adenovirus vector was extracted and identified.The transfection efficiency of the vector to HT-29 and LO2 was detected.The expression of Survivin mRNA and protein was detected by RT-PCR and Western blot.Results EGFP gene and shRNA gene targeting to human Survivin mRNA were inserted into conditionally replicating adenovirus vector successfully,proven by sequencing.The transfection efficiency to HT-29 of Ad-delE1b55kD-shRNA/Survivin-EGFP was significantly higher than replication defective adenovirus and liposome vector(P
Full text:
Available
Index:
WPRIM (Western Pacific)
Type of study:
Diagnostic study
Language:
Chinese
Journal:
Journal of Third Military Medical University
Year:
2003
Type:
Article
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