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Updated: Aug 20, 2026
Murine Model of CD40-activation of B cells
Published on: March 5, 2010
Construction of recombinant eukaryotic expression plasmid containing murine CD40 ligand gene and its expression in
Yong-Fang Jiang1, Yan He, Guo-Zhong Gong
1Center for Liver Diseases, Second Xiangya Hospital, Central South University, Changsha 410011, Hunan Province, China. jiangyongfang@hotmail.com
Aim:
To construct a recombinant murine CD40 ligand (mCD40L) eukaryotic expression vector for gene therapy and target therapy of hepatocellular carcinoma (HCC).
Methods:
mCD40L cDNA was synthesized by RT-PCR with the specific primers and directly cloned into T vector to generate middle recombinant. After digestion with restriction endonuclease, the target fragment was subcloned into the multi-clone sites of the eukaryotic vector. The constructed vector was verified by enzyme digestion and sequencing, and the product expressed was detected by RT-PCR and immunofluorescence methods.
Results:
The full-length mCD40L-cDNA was successfully cloned into the eukaryotic vector through electrophoresis, and mCD40L gene was integrated into the genome of infected H22 cells by RT-PCR. Murine CD40L antigen molecule was observed in the plasma of mCD40L-H22 by indirect immunofluorescence staining.
Conclusion:
The recombined mCD40L eukaryotic expression vector can be expressed in H22 cell line. It provides experimental data for gene therapy and target therapy of hepatocellular carcinoma.
