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Objective:The aim of the study was to construct miRNA-451 expression vector pLMP-miRNA-451 which could help identify the functions of miRNA-451 in SGC-7901 cel . Methods:Total RNA was extracted from SGC-7901 cel s to synthesized cDNA. The synthesized cDNA encoding pre-miRNA-451 was amplified by polymerase chain reaction (PCR). The PCR product was separated by electrophoresis on 1%agarose gel and then recovered and purified. The purified cDNA fragments of miRNA-451 precursor sequence was then ligated with vector pLMP for 1 h by using DNA ligase to form pLMP-miRNA-451 plasmid. After that, the pLMP-miRNA-451 plasmid was transformed into E. coli DH5αstrain expression system to clone and amplificate. The purified pLMP-miRNA-451 extracted from E. coli DH5αvia transformation and clone screening was identificatied with restriction enzyme digestion and DNA sequencing. At last, pLMP-miRNA-451 was transfected into SGC-7901 cel s with lip2000. Real-time PCR was used for detection of the miRNA-451, the transfection ef iciency was ob-served under fluorescence microscopy and cel counting kit-8 assay was conduced to evaluate the ef ect of miRNA-451 on SGC-7901 cel proliferation. Results:Our results showed that pLMP-miRNA-451 expression vector was not only constructed successful y and ef ectively infected SGC-7901 cel s, but also could repress the SGC-7901 cel proliferation. Conclusion:The constructed plasmid pLMP-miRNA-451 could used for further studies of miRNA-451 in SGC-7901 cel lines.
