Recombinant adenovirus of microRNA-193b inhibits proliferation of K562 cells
To construct a recombinant adenovirus vector containing human pre-miR-193b and investigate its effect on the proliferation of chronic myelocytic leukemia K562 cells. The cDNA of pre-miR-193b was obtained by chemical synthesis and inserted into the adenoviral shuttle vector pAdTrack-CMV. The recombin...
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Published in | Xi bao yu fen zi mian yi xue za zhi Vol. 31; no. 4; p. 452 |
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Main Authors | , , , , |
Format | Journal Article |
Language | Chinese |
Published |
China
01.04.2015
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Subjects | |
Online Access | Get more information |
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Summary: | To construct a recombinant adenovirus vector containing human pre-miR-193b and investigate its effect on the proliferation of chronic myelocytic leukemia K562 cells.
The cDNA of pre-miR-193b was obtained by chemical synthesis and inserted into the adenoviral shuttle vector pAdTrack-CMV. The recombinant shuttle plasmid was linearized by Pme I and transformed into AdEasier cells for homologous recombination in E.coli BJ5183 with the adenoviral backbone plasmid pAdEasy-1. The recombinant adenoviral plasmid was linearized by Pac I and then used for transfecting HEK293 cells. After package and amplification in HEK293 cells, the virus titer was determined by serial dilution assay. The expression level of miR-193b in K562 cells was detected by real-time quantitative PCR. The cell proliferation was observed by MTT assay.
The recombinant plasmid named pAd-miR-193b was confirmed by restriction enzyme analysis and sequencing. The recombinant adenovirus could infect K562 cells efficiently. Compared with control group, re |
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ISSN: | 1007-8738 |