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2006•Journal of Zhengzhou UniversityRequires access

Construction of hSAMP32 eukaryotic expression-vector and its expression in NIH3T3 cells

Hao Li

Open publisher page 1 citations

Abstract

Aim: To study the expression of hSAMP32 in NIH3T3 cells in vitro. Methods:To construct the pcDNA3.1(+)-hSAMP32 plasmid by PT-PCR and subcloning technique. The pcDNA3.1(+)-hSAMP32 DNA was transfected into NIH3T3 cells by liposome. The hSAMP32 mRNA expression was detected by in-situ hybridization.Results: RT-PCR product was consistent with that of target gene, hSAMP32. The target segment was detected through enzyme-digestion in subcloning.The bluish-violet colored granules were located in cytoplasm, but nothing was found in the cells transfected with pcDNA3.1 (+) or cells not transfected.Conclusion: The pcDNA3.1(+)-hSAMP32 DNA could be efficiently transfected into NIH3T3 cells by liposome-mediated gene transfection technique. The hSAMP32 gene expression in vitro is realized.

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What this paper is about

Aim: To study the expression of hSAMP32 in NIH3T3 cells in vitro. Methods:To construct the pcDNA3.1(+)-hSAMP32 plasmid by PT-PCR and subcloning technique. The pcDNA3.1(+)-hSAMP32 DNA was transfected into NIH3T3 cells by liposome. The hSAMP32 mRNA expression was detected by in-situ hybridization.Results: RT-PCR product was consistent with that of target gene, hSAMP32. The target segment was detected through enzyme-digestion in subcloning.The bluish-violet colored granules were located in cytoplasm, but nothing was found in the cells transfected with pcDNA3.1 (+) or cells not transfected.Conclusion: The pcDNA3.1(+)-hSAMP32 DNA could be efficiently transfected into NIH3T3 cells by liposome-mediated gene transfection technique. The hSAMP32 gene expression in vitro is realized.

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Available abstract

Aim: To study the expression of hSAMP32 in NIH3T3 cells in vitro. Methods:To construct the pcDNA3.1(+)-hSAMP32 plasmid by PT-PCR and subcloning technique. The pcDNA3.1(+)-hSAMP32 DNA was transfected into NIH3T3 cells by liposome. The hSAMP32 mRNA expression was detected by in-situ hybridization.Results: RT-PCR product was consistent with that of target gene, hSAMP32. The target segment was detected through enzyme-digestion in subcloning.The bluish-violet colored granules were located in cytoplasm, but nothing was found in the cells transfected with pcDNA3.1 (+) or cells not transfected.Conclusion: The pcDNA3.1(+)-hSAMP32 DNA could be efficiently transfected into NIH3T3 cells by liposome-mediated gene transfection technique. The hSAMP32 gene expression in vitro is realized.

Key concepts: Subcloning, Transfection, Molecular biology, In vitro, Plasmid, Gene, Gene expression, Cytoplasm

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