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Construction of Eukaryotic Expression Plasmid pcDNA3-FHIT and Its Transient Expression in COS-1 Cells 被引量:1

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摘要 Objective: To construct eukaryotic expression plasmid pcDNA3-fragile histidine triad(FHIT) and obtain its transient expression in COS-1 cells. Methods: FHIT gene was cloned from normal human thyroid tissue by RT-PCR and then inserted into eukaryotic expression vector pcDNA3. After the sequence was confirmed, the recombinant plasmid pcDNA3-FHIT was transfected into COS-1 cells by cation liposome. The transient expression in the cells was measured by immunocytochemistry. Results: The sequence of FHIT in pcDNA3 was correct and high expression was obtained in COS-1 cells. Conclusion: The eukaryotic expression plasmid pcDNA3-FHIT was constructed successfully and could highly express FHIT protein in COS-1 cells. This will be potentially useful for the research on gene therapy.
出处 《Journal of Nanjing Medical University》 2004年第5期275-277,共3页 南京医科大学学报(英文版)
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  • 1田中伟,冯捷,宋向凤,马慧群,彭振辉.皮肤鳞状细胞癌中FHIT蛋白的检测及其临床意义[J].中国皮肤性病学杂志,2005,19(2):68-70. 被引量:7
  • 2Ohta M, Inoue H, Cotticelli MG, et al. The FHIT gene, spanning the chromosome 3p14. 2 fragile site and renal carcinoma-associated t(3 ;8) breakpoint, is abnormal in digestive tract cancers [ J]. Cell, 1996,84:587 - 597.
  • 3Deng WG, Nishizaki M, Fang B, et al. Induction of apoptosis by tumor suppressor FHIT via death receptor signaling pathway in human lung cancer cells [ J ]. Biochem Biophys Res Commun, 2007, 355:993 - 999.
  • 4Guo Z, Vishwanatha JK. Effect of regulated expression of the fragile histidine triad gene on cell cycle and proliferation [J]. Mol Cell Bioehem, 2000, 204(122) :83 -88.
  • 5Ishii H, Dumon KR, Vecchione A, et al. Effect of adenoviral transduction of the fragile histidine triad gene into esophageal cancer cells [ J ]. Cancer Res, 2001, 61 (4) : 1578 - 1584.
  • 6Pichiorri F, Trapasso F, Palumbo T, et al. Preclinical assessment of FHIT gene replacement therapy in human leukemia using a chimeric adenovirus, AdS/F35 [ J ]. Clin Cancer Res, 2006, 12(1) :3494-3501.
  • 7Nakagawa Y, Akao Y. FHIT protein inhibits cell growth by attenuating the signaling mediated by nuclear factor-κB in colon cancer cell lines [ J]. Exp Cell Res, 2006, 312 (13) :2433 - 2442.
  • 8Dumon KR, Ishii H, Fong LY, prevents tumor development in et al. FHIT gene therapy FHIT-deficient mice [ J ]. Proe Natl Acad Sci USA, 2001, 98(6) :3346 -3351

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