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Published on: July 12, 2013
[Construction and identification of human PRMT1 gene eukaryotic vector]
Chen-Yan Li1, Qing-Zhu Sun, Xu-Dong Yang
1Department of Medical Genetics and Molecular Biology, Xi'an Jiaotong University College of Medicine, Xi'an 710061, China.
Summary
Researchers successfully constructed the pcDNA3.1(+)-PRMT1 expression plasmid. This recombinant plasmid efficiently expresses PRMT1 (Protein arginine methyltransferase 1) in A549 cells, validated by molecular assays.
Area of Science:
- Molecular Biology
- Gene Expression
- Recombinant DNA Technology
Background:
- Protein arginine methyltransferase 1 (PRMT1) plays crucial roles in cellular processes.
- Efficient expression systems are vital for studying PRMT1 function and regulation.
- The pcDNA3.1(+) vector is a common tool for eukaryotic gene expression.
Purpose of the Study:
- To construct a eukaryotic recombinant expression plasmid for human PRMT1.
- To validate the efficient expression of PRMT1 from the constructed plasmid in cultured cells.
Main Methods:
- Human PRMT1 cDNA was amplified using reverse transcription polymerase chain reaction (RT-PCR).
- The amplified PRMT1 gene was cloned into the pcDNA3.1(+) vector.
- Transfection into A549 cells followed by Real-time PCR and Western blot analysis for PRMT1 expression.
Main Results:
- The pcDNA3.1(+)-PRMT1 recombinant plasmid was successfully constructed and verified by sequencing.
- The inserted PRMT1 sequence matched the NCBI GenBank reference.
- Efficient mRNA and protein expression of PRMT1 was confirmed in transfected A549 cells.
Conclusions:
- The pcDNA3.1(+)-PRMT1 recombinant expression plasmid was successfully constructed.
- This construct serves as an effective tool for studying PRMT1 in eukaryotic systems.
