Green Fluorescent Protein Retroviral Vectors: Low Titer and High Recombination Frequency Suggest a Selective Disadvantage
作者： Yutaka Hanazono , Jian-Mei Yu , Cynthia E. Dunbar , Robert V. B. Emmons
DOI: 10.1089/HUM.1997.8.11-1313
关键词: Green fluorescent protein 、 Gene expression 、 Vector (molecular biology) 、 Cell culture 、 Gene 、 Transfection 、 Molecular biology 、 Biology 、 Mutant 、 Plasmid
摘要: ABSTRACT Green fluorescent protein (GFP) has been used as a reporter molecule for gene expression because it fluoresces green after blue-light excitation. Inclusion of this in vector could allow rapid, nontoxic selection successfully transduced cells. However, many attempts by our laboratory to isolate stable retroviral producer cell clones secreting biologically active vectors containing either the highly S65T-GFP mutant or humanized GFP have failed. Vector plasmids various forms and neomycin resistance were transfected into three different packaging lines fluorescence was observed several days, but selected with G418 no longer fluoresced. Using confocal microscopy, brightest cells contract die within matter days. RNA slot-blot analysis supernatants showed viral production from plasmid-transfected clones, although all derived transfection an identic...
liebertonline.com参考文章(15)
Jim Haseloff, Brad Amos, GFP in plants. Trends in Genetics. ,vol. 11, pp. 328- 329 ,(1995) , 10.1016/0168-9525(95)90186-8
R. K. Humphries, S. Karlsson, M. Migita, J. A. Medin, R. Pawliuk, S. Jacobson, J. W. Nagle, S. Anderson, M. Amiri, SELECTION OF TRANSDUCED CD34+ PROGENITORS AND ENZYMATIC CORRECTION OF CELLS FROM GAUCHER PATIENTS, WITH BICISTRONIC VECTORS Proceedings of the National Academy of Sciences of the United States of America. ,vol. 92, pp. 12075- 12079 ,(1995) , 10.1073/PNAS.92.26.12075
Roger Heim, Andrew B. Cubitt, Roger Y. Tsien, Improved green fluorescence Nature. ,vol. 373, pp. 663- 664 ,(1995) , 10.1038/373663B0
Douglas C. Prasher, Virginia K. Eckenrode, William W. Ward, Frank G. Prendergast, Milton J. Cormier, Primary structure of the Aequorea victoria green-fluorescent protein. Gene. ,vol. 111, pp. 229- 233 ,(1992) , 10.1016/0378-1119(92)90691-H
W. S. Pear, G. P. Nolan, M. L. Scott, D. Baltimore, Production of high titer helper-free retroviruses by transient transfection Proceedings of the National Academy of Sciences of the United States of America. ,vol. 90, pp. 8392- 8396 ,(1994) , 10.1073/PNAS.90.18.8392
M Chalfie, Y Tu, G Euskirchen, W. Ward, D. Prasher, Green fluorescent protein as a marker for gene expression Science. ,vol. 263, pp. 802- 805 ,(1994) , 10.1126/SCIENCE.8303295
Joseph Alper, Genomica set to score twice Nature Biotechnology. ,vol. 16, pp. 610- 610 ,(1998) , 10.1038/NBT0798-610B
Cynthia E. Dunbar, Robert V. B. Emmons, Gene transfer into hematopoietic progenitor and stem cells: progress and problems. Stem Cells. ,vol. 12, pp. 563- 576 ,(1994) , 10.1002/STEM.5530120604
Steven F. Ziegler, Cheryl A. Whitlock, Steven P. Goff, Ann Gifford, Owen N. Witte, Lethal effect of the abelson murine leukemia virus transforming gene product Cell. ,vol. 27, pp. 477- 486 ,(1981) , 10.1016/0092-8674(81)90389-5
Linzhao Cheng, Joan Fu, Ann Tsukamoto, Robert G Hawley, None, Use of green fluorescent protein variants to monitor gene transfer and expression in mammalian cells Nature Biotechnology. ,vol. 14, pp. 606- 609 ,(1996) , 10.1038/NBT0596-606