Related Experiment Video
Updated: Jul 19, 2026
A Convenient and General Expression Platform for the Production of Secreted Proteins from Human Cells
Published on: July 31, 2012
[Construction and expression of the eukaryotic plasmids containing different subunits of human IL-12 gene]
Mu Hao1, Lang Bao, Hui-Dong Zhang
1Department of Infection and Immunity, Preclinical and Forensic Medicine School of West China Medical Center, Sichuan University, Chengdu 610041, China. haomusg@163.com
Aim:
To clone and construct the eukaryotic expression plasmids containing different subunits of human IL-12 and to investigate its expression following transient transfection into COS-7 cells.
Methods:
Human leukaemia cells (HL-60 and THP-1) were stimulated by DMSO, IFN-gamma and LPS, and the target genes including IL-12 p40 and p35 subunits were amplified by RT-PCR. P40 and p35 fragments were then connected and the recombinant plasmids pcDNA-p40, pcDNA-p35 and pcDNA-p40-p35 were constructed by double restriction enzymes digestion and connection. Positive plasmids were transiently transfected into COS-7 cells with siPORT XP-1 reagent. The expression of target gene was detected by RT-PCR and ELISA.
Results:
The recombinant plasmids were confirmed by PCR amplification, double restriction enzymes digestion and DNA sequencing. Target gene expression was detected by RT-PCR and ELISA in the transfected COS-7 cells.
Conclusion:
The successful construction of the recombinant plasmids pcDNA-p40, pcDNA-p35 and pcDNA-p40-p35 provides a useful tool for further research on the immune regulatory role of IL-12.
More Related Videos
10:06Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells
Published on: April 26, 2017
10:18Generation of Plasmid Vectors Expressing FLAG-tagged Proteins Under the Regulation of Human Elongation Factor-1α Promoter Using Gibson Assembly
Published on: February 9, 2015