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荧光定量PCR检测Kbpala/Alb-ATP7B短暂转染BRL细胞后的转录情况

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摘要 【目的】明确 Kbpala/Alb-ATP7B 短暂转染 BRL 细胞后的表达情况。【方法】将 Kbpala/Alb-ATP7B 用阳离子脂质体 Lipofect AMINE^(TM)2000 Reagent 转染 BRL 细胞后,按转染后不同时间点提取细胞 mRNA,逆转录后,使用 SYBR Green 荧光定量 PCR 检测其表达情况。【结果】转染后4h 即可检测到外源基因的表达,48h 外源基因的表达最高,可持续7d 以上。【结论】Kbpala/Alb-ATP7B 可在 BRL 细胞中有效表达,表明载体构建成功。
出处 《中山大学学报(医学科学版)》 CAS CSCD 北大核心 2005年第B03期56-58,共3页 Journal of Sun Yat-Sen University:Medical Sciences
基金 211工程重点学科建设课题(98138) 广东省自然科学基金(990064) 卫生部临床学科重大项目(2001321)
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  • 1石铸,王莹,任廷文,梁秀龄,徐评议,李洵桦,邱国光.TX小鼠的铜代谢特点和肝脏病理学特征[J].中山大学学报(医学科学版),2003,24(3):211-213. 被引量:11
  • 2徐琳,梁秀龄,徐评议,王尚武,朱雅南,丰岩清,谢秋幼.小鼠肝脏特异性表达载体ALB-ATP7B的构建[J].中山大学学报(医学科学版),2004,25(1):34-38. 被引量:4
  • 3Izban MG, Papaeonstantinou J. Cell-specific expression of mouse albumin promoter. Evidence for cell-specific DNA elements within the proximal promoter region and cis-acting DNA elements upstream of -160[J]. J Biol Chem, 1989, 264 (16): 9171-9.
  • 4Wu GY, Wilson JM, Shalaby F, et al. Receptor-mediated gene delivery in vivo. Partial correction of genetic analbuminemia in Nagase rats[J]. J Biol Chem, 1991, 266(22): 14338-42.
  • 5Ralph WM Jr, Cabatingan MS, Schaffer PA. Induction of herpes simplex virus type 1 immediate-early gene expression by a cellular activity expressed in Vero and NB41A3 cells after growth arrest-release[J]. J Virol, 1994, 68(11):6871-82.
  • 6Adamson R, Lyons K, Sharrard M, et al. Transient transfection of Theileria annulata[J]. Mol Biochem Parasitol, 2001, 114(1): 53-61.
  • 7蒋欣泉,张志愿,曹俊,陈传俊,王明国,周晓健,陈万涛.绿色荧光蛋白基因转染骨髓基质干细胞的瞬时表达检测[J].华西口腔医学杂志,2003,21(2):89-91. 被引量:6
  • 8Skovhus TL, Ramsing NB, Holmstrom C,et al Real-time quantitative PCR for assessment of abundance of pseudoalteromonas species in marine samples[J]. Appl Environ Microbiol, 2004, 70(4):2373-82.
  • 9Morrison TB, Weis J J, Wittwer CT. Quantification of low-copy transcripts by continuous SYBR Green Ⅰ monitoring during amplification[J]. Biotechniques, 1998, 24(6):954-8.
  • 10Yin JL, Shackel NA, Zekry A. Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) for measurement of cytokine and growth factor mRNA expression with fluorogenic probes or SYBR Green 1 [J]. Immunol Cell Biol, 2001, 79(3):213-21.

二级参考文献15

  • 1[1]Prockop DJ. Marrow stromal cells as stem cells for nonhematopoietic tissues. Science,1997,276(5309):71-74
  • 2[2]Chalfie M, Tu Y, Euskirchen G, et al. Green fluorescent protein as a marker for gene expression. Science,1994,263(5148):802-805
  • 3[3]Cormack BP, Valdivia RH, Falkow S. FACS-optimized mutants of the green fluorescent protein (GFP). Gene,1996,173(1):33-38
  • 4[4]Evans CH, Robbins PD. Genetically augmented tissue engineering of the musculoskeletal system. Clin Orthop,1999,367(suppl):410-418
  • 5[5]Moutsatsos IK, Turgeman G, Zhou S,et al. Exogenously regulated stem cell-mediated gene therapy for bone regeneration. Mol Ther, 2001,3(4):449-461
  • 6Theophilos M B, Cox D W, Mercer J F.The toxic milk mouse is a murine model of Wilson disease[J].Hum Mol Genet, 1996, 5 (10): 1619.
  • 7Roelofsen H, Wolters H, Vanluyn M, et al.Copper- induced apical trafficking of ATP7B in polarized hepatoma cells provides a mechanism for biliary copper excretion[J].Gastroenterology, 2000, 119(3):782.
  • 8Schaefer M, Roelofsen H, Wolters H, et al.Localization of the Wilson's disease protein in human liver[J].Gastroenterology, 1999, 117(6):1380.
  • 9Mercer J F.The molecular basis of copper- transport diseases[J].Trends Mol Med, 2001, 7(2): 64.
  • 10Strain A J,Crosby H A.Hepatic stem cells[J].Gut, 2000, 46(6):743.

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