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.
AIM: To construct a recombinant murineCD40 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. MurineCD40L 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.
Authors: Clemens-Martin Wendtner; David M Kofler; Hans D Theiss; Christian Kurzeder; Raymund Buhmann; Carmen Schweighofer; Luca Perabo; Susanne Danhauser-Riedl; Jens Baumert; Wolfgang Hiddemann; Michael Hallek; Hildegard Büning Journal: Blood Date: 2002-09-01 Impact factor: 22.113
Authors: E M Sotomayor; I Borrello; E Tubb; F M Rattis; H Bien; Z Lu; S Fein; S Schoenberger; H I Levitsky Journal: Nat Med Date: 1999-07 Impact factor: 53.440
Authors: F Mach; U Schönbeck; G K Sukhova; T Bourcier; J Y Bonnefoy; J S Pober; P Libby Journal: Proc Natl Acad Sci U S A Date: 1997-03-04 Impact factor: 11.205