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Construction and expressin of eukaryotic expression vector pcDNA3.0-Cx43 in mesenchymal stem cells

He Zhou

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Abstract

AIM: To construct a eukaryotic expression vector encoding full length connexin43 (Cx43) cDNA and to investigate its overexpression in mesenchymal stem cells (MSCs). METHODS: Full length Cx43 cDNA was amplified in E. coli JM109 and cloned into the eukaryotic expression vector pcDNA3.0. Stably transfected cell colonies were obtained by gene transfer and G418 screening and control cells were transfected with pcDNA3.0 and screened by using the same method. RESULTS: Electrophoresis results indicated that the recombinant vector was constructed correctly. Immunocytochemical analysis revealed an overexpression of Cx43 in pcDNA3.0 Cx43 transfected MSCs. CONCLUSION: The connexin43 can be expressed in MSCs.

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What this paper is about

AIM: To construct a eukaryotic expression vector encoding full length connexin43 (Cx43) cDNA and to investigate its overexpression in mesenchymal stem cells (MSCs). METHODS: Full length Cx43 cDNA was amplified in E. coli JM109 and cloned into the eukaryotic expression vector pcDNA3.0. Stably transfected cell colonies were obtained by gene transfer and G418 screening and control cells were transfected with pcDNA3.0 and screened by using the same method. RESULTS: Electrophoresis results indicated that the recombinant vector was constructed correctly. Immunocytochemical analysis revealed an overexpression of Cx43 in pcDNA3.0 Cx43 transfected MSCs. CONCLUSION: The connexin43 can be expressed in MSCs.

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Available abstract

AIM: To construct a eukaryotic expression vector encoding full length connexin43 (Cx43) cDNA and to investigate its overexpression in mesenchymal stem cells (MSCs). METHODS: Full length Cx43 cDNA was amplified in E. coli JM109 and cloned into the eukaryotic expression vector pcDNA3.0. Stably transfected cell colonies were obtained by gene transfer and G418 screening and control cells were transfected with pcDNA3.0 and screened by using the same method. RESULTS: Electrophoresis results indicated that the recombinant vector was constructed correctly. Immunocytochemical analysis revealed an overexpression of Cx43 in pcDNA3.0 Cx43 transfected MSCs. CONCLUSION: The connexin43 can be expressed in MSCs.

Key concepts: Transfection, Complementary DNA, Mesenchymal stem cell, Molecular biology, Recombinant DNA, Vector (molecular biology), Biology, Gene

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