Construction of recombinant human SMOC1 gene adenovirus vector and its expression in human aortic valve interstitial cells
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    Abstract:

    Objective:To construct a recombinant adenovirus vector containing SMOC1 gene and to identify its expression in cultured human aortic valve interstitial cells (hAVICs) and effects on cell proliferation. Methods:The SMOC1 gene was amplified by PCR and inserted into pGEM-T Easy vector and sequenced. Double digested with Not I and Xho I,SMOC1 gene was inserted into pShuttle-CMV vector. This recombinant plasmid was linearized by Pme I and electronically transfected into BJ5183 germs to get the recombinant adenovirus vector (pAdEasy-SMOC1). Then the recombinant adenovirus expressing SMOC1 (Ad-SMOC1) was obtained by packaging in HEK293 A cells. After infecting hAVICs by Ad-SMOC1,the expression of recombinant SMOC1 was detected by Western blot and Immunofluorescence. The proliferation of hAVICs infected with Ad-SMOC1 was measured by MTS assay. Results:The recombinant adenovirus Ad-SMOC1 was successfully generated,which could effectively infect hAVICs and increase SMOC1 protein levels in hAVICs. Besides this,exogenous expressed SMOC1 was mainly located at the cytoplasm and membrane of hAVICs. MTS assay showed that recombinant SMOC1 promoted the proliferation of hAVICs. Conclusion:The bioactive recombinant adenovirus Ad-SMOC1 has been successfully constructed and recombinant SMOC1 promotes cell proliferation. This study laid a foundation for further research of the function of SMOC1 in hAVICs.

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赵 蓉,王 惠,杨 杨,李名鹏,孔祥清,孙 伟.人SMOC1重组腺病毒载体的构建及其在主动脉瓣间质细胞中的表达[J].南京医科大学学报(自然科学版英文版),2012,(12):1635-1640.

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  • Online: January 09,2013
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