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Published on: July 7, 2013
[The effect of pcDNA 3.1/RPs15 on skin fibroblasts in vitro]
Sheng-jian Tang1, Xiao-ke Wang, Xiao-jie LV
1Institute of Plastic Surgery, Weifang Medical University, Weifang 261042, China.
Objective:
To construct eukaryotic expression vector of ribosomal protein sl5(RPs15) gene and study its effect on mouse skin fibroblasts in vitro.
Methods:
The RPs15 cDNA encoding region of fetal mouse skin was amplified by reverse transcription-polymerase chain reaction (RT-PCR) method and cloned into eukaryotic expression vector pcDNA3.1(-). The recombinant plasmid was transfected into adult mouse skin fibroblasts by FuGENE6 transfection reagents. Then the expression of RPs15 gene, was detected and its biological effect on fibroblasts was measured.
Results:
The DNA sequencing result of pcDNA3.1/RPs15 was identical with the reported. The RPs15 gene was expressed in transfected fibroblasts. The growth density of fibroblasts decreased with the conformation changing accordingly.
Conclusions:
The eukaryotic expression vector pcDNA3.1/RPs15 is successively constructed and can be expressed in mouse skin fibroblasts. The results set up a basis for further study of the effect of RPs15 gene on skin fibroblasts.
Insights
Researchers constructed a ribosomal protein S15 (RPs15) gene expression vector and confirmed its expression in mouse skin fibroblasts. This vector successfully altered fibroblast growth and conformation, providing a basis for further research.
Area of Science:
- Molecular Biology
- Cell Biology
Context:
- Ribosomal proteins play crucial roles in protein synthesis and cellular functions.
- Understanding the specific roles of individual ribosomal proteins, like RPs15, in mammalian cells is essential.
Purpose:
- To construct a eukaryotic expression vector for the ribosomal protein S15 (RPs15) gene.
- To investigate the in vitro effects of RPs15 gene expression on mouse skin fibroblasts.
Summary:
- The RPs15 cDNA was amplified and cloned into the pcDNA3.1(-) vector, creating the recombinant plasmid pcDNA3.1/RPs15.
- This plasmid was transfected into adult mouse skin fibroblasts, and RPs15 gene expression was confirmed via sequencing and detection.
- Transfected fibroblasts exhibited decreased growth density and altered conformation.
Impact:
- Successfully constructed and validated a functional eukaryotic expression vector for RPs15.
- Demonstrated that RPs15 expression influences mouse skin fibroblast proliferation and morphology.
- Provides a foundational tool for further studies on RPs15's role in skin fibroblast biology and potential therapeutic applications.
