Variables Controlling the Expression Level of Exogenous Genes in Dictyostelium
作者： Ka Ming Pang , Michael A. Lynes , David A. Knecht
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摘要: Ectopic expression of genes from recombinant plasmids is commonly used to study gene function. In Dictyostelium, three drug resistance cassettes are as selectable markers in vectors. We report here a comparative the green fluorescent protein (GFP) vectors containing each drug-resistant cassettes. The was highest cells transformed with neomycin-resistant cassette (pDNeoGFP), followed by hygromycin-resistant (pDHygGFP) and blasticidin-resistant (pDBsrGFP). level GFP directly related copy number vector transformants. turn, depended on well concentration selection. general, higher numbers could be selected concentration. also affected method transformation. For pDHygGFP, much electroporation than those calcium phosphate coprecipitation. However, only slight difference observed for pDNeoGFP or pDBsrGFP.
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sci-hub.se 参考文章(15)
B. Reiss, R. Sprengel, H. Schaller, Protein fusions with the kanamycin resistance gene from transposon Tn5. The EMBO Journal. ,vol. 3, pp. 3317- 3322 ,(1984) , 10.1002/J.1460-2075.1984.TB02297.X
T T Egelhoff, S S Brown, D J Manstein, J A Spudich, Hygromycin resistance as a selectable marker in Dictyostelium discoideum. Molecular and Cellular Biology. ,vol. 9, pp. 1965- 1968 ,(1989) , 10.1128/MCB.9.5.1965
W Nellen, C Silan, R A Firtel, DNA-mediated transformation in Dictyostelium discoideum: regulated expression of an actin gene fusion. Molecular and Cellular Biology. ,vol. 4, pp. 2890- 2898 ,(1984) , 10.1128/MCB.4.12.2890
D A Knecht, S M Cohen, W F Loomis, H F Lodish, Developmental regulation of Dictyostelium discoideum actin gene fusions carried on low-copy and high-copy transformation vectors. Molecular and Cellular Biology. ,vol. 6, pp. 3973- 3983 ,(1986) , 10.1128/MCB.6.11.3973
Dietmar J. Manstein, Hans-Peter Schuster, Piero Morandini, Deborah M. Hunt, Cloning vectors for the production of proteins in Dictyostelium discoideum Gene. ,vol. 162, pp. 129- 134 ,(1995) , 10.1016/0378-1119(95)00351-6
H. Adachi, T. Hasebe, K. Yoshinaga, T. Ohta, K. Sutoh, Isolation of Dictyostelium discoideum cytokinesis mutants by restriction enzyme-mediated integration of the blasticidin S resistance marker. Biochemical and Biophysical Research Communications. ,vol. 205, pp. 1808- 1814 ,(1994) , 10.1006/BBRC.1994.2880
C. Barth, D.J. Fraser, P.R. Fisher, Co-insertional replication is responsible for tandem multimer formation during plasmid integration into the Dictyostelium genome. Plasmid. ,vol. 39, pp. 141- 153 ,(1998) , 10.1006/PLAS.1997.1326
Wolfgang Nellen, Richard A. Firtel, High-copy-number transformants and co-transformation in Dictyostelium. Gene. ,vol. 39, pp. 155- 163 ,(1985) , 10.1016/0378-1119(85)90309-9
Kazuo Sutoh, A Transformation Vector for Dictyostelium discoideum with a New Selectable Marker bsr Plasmid. ,vol. 30, pp. 150- 154 ,(1993) , 10.1006/PLAS.1993.1042
Eunkyung Lee, Eric A. Shelden, David A. Knecht, Formation of F-Actin Aggregates in Cells Treated With Actin Stabilizing Drugs Cytoskeleton. ,vol. 39, pp. 122- 133 ,(1998) , 10.1002/(SICI)1097-0169(1998)39:2<122::AID-CM3>3.0.CO;2-8