Intramolecular integration assay validates integrase phi C31 and R4 potential in a variety of insect cells.
Phage phi C31 and R4 integrases are site-specific and unidirectional serine recombinases. We have analyzed the ability of these integrases to mediate intramolecular integration between their attB and attP sites in 7 important insect cell lines as a means of predicting their relative mobility in the corresponding insect species. Both integrases exhibit significantly higher frequencies in Drosophila S2 cells than in the other insect cell lines examined, but do work well in all of the species tested. Our results, coupled with previous results of the activity of phi C31 integrase in D. melanogaster and Aedes aegypti, suggest the family of serine catalyzed integrases will be useful site-specific integration tools for functional genome analysis and genetic engineering in a wide range of insect species.
Chompoosri
Jakkrawarn
J
Department of Biological Sciences, Eck Institute of Global Health, University of Notre Dame, Notre Dame, IN 46556, USA.
Fraser
Tresa
T
Rongsriyam
Yupha
Y
Komalamisra
Narumon
N
Siriyasatien
Padet
P
Thavara
Usavadee
U
Tawatsin
Apiwat
A
Fraser
Malcolm J
MJ
D000330:Aedes / Q000201:enzymology / Q000235:genetics*; D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D001287:Attachment Sites, Microbiological; D001435:Bacteriophages / Q000201:enzymology* / Q000235:genetics; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D017931:DNA Primers; D004330:Drosophila / Q000201:enzymology / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011995:Recombination, Genetic / Q000235:genetics; D014162:Transfection
D017931:DNA Primers; D008156:Luciferases; D019426:Integrases