JI Yewei, et al. Construction of si-RNA synthesis plasmids targeting human Hsp90β gene and observation of their transfection efficiencies. [J]. Journal of Chongqing Medical University (1):8-11(2007)
JI Yewei, et al. Construction of si-RNA synthesis plasmids targeting human Hsp90β gene and observation of their transfection efficiencies. [J]. Journal of Chongqing Medical University (1):8-11(2007)DOI:
Construction of si-RNA synthesis plasmids targeting human Hsp90β gene and observation of their transfection efficiencies
Objective:To construct the plasmid that can direct the synthesis of si-RNA like transcripts to specifically and effectively in hibit the mRNA level of human Hsp90βgene
and to compare the transfection efficiencies of the plasmids in different cell strains. Methods:Three 64 nt oligos corresponding to different regions in the target gene are chemically synthesized and annealed
then inserted into pSUPER EGFP1
proliferated by DH5a
and determined by endodigestion and sequencing. Three strains
HepG2、HUVEC and HeK293
were cultured. We then transfected the plasmids into the cells under different ratio of plasmid and Lipofectamine
and compared the transfection efficiencies of them by detection of EGFP Fluoresence. Results:The presence of positive recombinant clones was verified by double-digestion and sequencing. The bases inserted into the pasmids were correct.And the positive colonies were named as pSuper-Hsp90β1、pSuper-Hsp90β2 and pSuper-Hsp90β3. and the same condition
the constructed plasmid had the highest transfection efficiency in HeK293
and the lowest in HepG2 cells. Conclusions:The si-RNA synthesis plasmids targeting Hsp90β could be successfully constructed by using psuper plasmids
and the difference of their transfection efficiencies in different cells was significant.
Molecular mechanism of Buzhong Yiqi Decoction in coordination with Siβ-catenin in intervention against cisplatin resistance in lung adenocarcinoma by regulating the Wnt/β-catenin signaling pathway
The optimal electroporation condition of recombinant plasmid
Effect of silencing mdrl gene by RNA interference with adriamycin on proliferation of SKOV3/ADM
Construction of pEGFP-VEGF recombinant plasmid and its expression in osteoblast
Construction and verification of eukaryotic expression vector for mouse Hsp84
Related Author
Gao Yuan
Yu Dan
Wang Ying
Wang Zhe
Mu Qirui
Li He
任红
et al
Related Institution
Department of Pathology,College of Integrative Medicine,Liaoning University of Traditional Chinese Medicine
重庆医科大学病毒性肝炎研究所
Department of Obstetrics and Gynecology,the Second College of Clinical Medicine,Chongqing Medical University
重庆医科大学临床学院骨科
Department of Orthopaedics, College of Clinical Medicine , Chongqing Medical University