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Cloning of chicken anemia virus vp3 gene and apoptosis inductive effect of vp3 gene in vitro / 华中科技大学学报(医学)(英德文版)
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 329-334, 2003.
Article in English | WPRIM | ID: wpr-330914
ABSTRACT
Using PCR technique, the vp3 gene of chicken anemia virus (CAV) was cloned into the eukaryotic expression vector pcDNA3 to construct a recombinant pcDNA-vp3. Restriction enzyme digestion and sequencing analysis revealed that CAV vp3 gene was correctly inserted into the blank vector pcDNA3. After LipofectAMINE-mediated transfection in vitro with pcDNA-vp3 and pcDNA3 respectively, the total mRNA was extracted from liver carcinoma cell lines HepG2 and diploid cell line L-02, and RT-PCR was performed afterward. The results of RT-PCR suggested that vp3 gene was expressed in these two cell lines. At the same time, using in situ apoptotic detection assay, TUNEL kits, the apoptotic cells were found in pcDNA-vp3 transfected HepG2, but not in mock transfected cell lines. VP3 could induce cell death by apoptosis in cancer cell lines, but not in diploid cell lines. All the results indicated that CAV vp3 gene, a potential therapeutic agents, has the potential of being used for cancer treatment.
Subject(s)
Full text: Available Index: WPRIM (Western Pacific) Main subject: Pathology / Pharmacology / Viral Proteins / Recombinant Proteins / Transfection / Chickens / Polymerase Chain Reaction / Cloning, Molecular / Sequence Analysis, DNA / Apoptosis Limits: Animals / Humans Language: English Journal: Journal of Huazhong University of Science and Technology (Medical Sciences) Year: 2003 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Pathology / Pharmacology / Viral Proteins / Recombinant Proteins / Transfection / Chickens / Polymerase Chain Reaction / Cloning, Molecular / Sequence Analysis, DNA / Apoptosis Limits: Animals / Humans Language: English Journal: Journal of Huazhong University of Science and Technology (Medical Sciences) Year: 2003 Type: Article