Reference Report

Reference
Citation	Groth, A.C., Fish, M., Nusse, R., Calos, M.P. (2004). Construction of Transgenic Drosophila by Using the Site-Specific Integrase From Phage {phi}C31. Genetics 166(4): 1775--1782. (Export to RIS)
FlyBase ID	FBrf0174693
Publication Type	Research paper
PubMed ID	15126397
PubMed Abstract	The phiC31 integrase functions efficiently in vitro and in Escherichia coli, yeast, and mammalian cells, mediating unidirectional site-specific recombination between its attB and attP recognition sites. Here we show that this site-specific integration system also functions efficiently in Drosophila melanogaster in cultured cells and in embryos. Intramolecular recombination in S2 cells on transfected plasmid DNA carrying the attB and attP recognition sites occurred at a frequency of 47%. In addition, several endogenous pseudo attP sites were identified in the fly genome that were recognized by the integrase and used as substrates for integration in S2 cells. Two lines of Drosophila were created by integrating an attP site into the genome with a P element. phiC31 integrase injected into embryos as mRNA functioned to promote integration of an attB-containing plasmid into the attP site, resulting in up to 55% of fertile adults producing transgenic offspring. A total of 100% of these progeny carried a precise integration event at the genomic attP site. These experiments demonstrate the potential for precise genetic engineering of the Drosophila genome with the phiC31 integrase system and will likely benefit research in Drosophila and other insects.
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Language of Publication	English
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Publication Type	Journal
Abbreviation	Genetics
Title	Genetics
Publication Year	1916-
ISBN/ISSN	0016-6731
Data from Reference
Alleles (5)
	Ecol\lacZ^BCPB+ phiC31\int^cGa phiC31\int^MtnA.PG w^+mC y^+t*
Constructs (6)
	P{CaryP} P{UASTB} pET11φC31 pET11φC31poly(A) pP{CaryP} pP{UASTB}
Genes (12)
	attP:Dps1 attP:Dps2 attP:Dps3 CG6310 Ecol\lacZ Mocs1 phiC31\attP phiC31\int sbb Sliv\attB w y
Insertions (2)
	P{CaryP}attP1 P{CaryP}attP2