The Tol2kit: a multisite gateway-based construction kit for Tol2 transposon transgenesis constructs.
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"The Tol2kit: a multisite gateway-ba..." refers methods in this paper
...Embryos were raised at 28.5°C and staged according to time postfertilization and morphology ( Kimmel et al., 1995 )....
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...Embryos were raised at 28.5°C and staged according to time postfertilization and morphology (Kimmel et al., 1995)....
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4,607 citations
"The Tol2kit: a multisite gateway-ba..." refers background in this paper
...The two fluorescent proteins used were EGFP (Zhang et al., 1996) and the monomeric red fluorescent protein mCherry ( Shaner et al., 2004; Gray et al., 2006)....
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...…have not been able to visualize the mCherry protein, likely for three reasons: the relative dimness of mCherry compared with EGFP, the low expression level driven by the IRES, and suboptimal imaging using 543-nm excitation, far from the peak of mCherry excitation at 587 nm (Shaner et al., 2004)....
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...We have also generated a set of equivalent IRES constructs driving mCherry, but have not been able to visualize the mCherry protein, likely for three reasons: the relative dimness of mCherry compared with EGFP, the low expression level driven by the IRES, and suboptimal imaging using 543-nm excitation, far from the peak of mCherry excitation at 587 nm ( Shaner et al., 2004 )....
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...The two fluorescent proteins used were EGFP (Zhang et al., 1996) and the monomeric red fluorescent protein mCherry (Shaner et al., 2004; Gray et al., 2006)....
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1,401 citations
"The Tol2kit: a multisite gateway-ba..." refers background in this paper
...Three of the 3 entry clones encode such tags in the standard Gateway reading frame: p3E-MTpA encodes a 6 myc epitope tag ( Munro and Pelham, 1986; Roth et al., 1991) derived from pCS2MT; p3E-EGF-PpA encodes an EGFP tag; and p3EmCherrypA encodes an mCherry tag....
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1,312 citations
"The Tol2kit: a multisite gateway-ba..." refers background in this paper
...The final middle entry clone encodes Gal4VP16, a fusion of the Gal4 DNA binding domain to the highly acidic region of the herpes simplex virus protein VP16, which strongly activates transcription downstream of the Gal4 UAS ( Sadowski et al., 1988 )....
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...The Fse-Asc cassette was generated by annealing DNA oligos, and the UAS insert was conventionally cloned from pBUAS-E1b-RFP (gift of R. Köster)....
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...The final middle entry clone encodes Gal4VP16, a fusion of the Gal4 DNA binding domain to the highly acidic region of the herpes simplex virus protein VP16, which strongly activates transcription downstream of the Gal4 UAS (Sadowski et al., 1988)....
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...The p5E-UAS contains a 10 UAS multimerized Gal4 upstream activating sequence element from yeast, followed by an adenovirus E1b TATA box and a carp beta-actin 5 -UTR fragment (Rorth, 1996; Köster and Fraser, 2001)....
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...This cassette is designed to be a transgenesis marker, particularly useful when the transgene of interest is nonfluorescent, difficult to visualize (e.g., a few neurons in the brain), or conditionally expressed (e.g., driven by a UAS sequence or a heatshock promoter)....
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1,085 citations
"The Tol2kit: a multisite gateway-ba..." refers methods in this paper
...First, recombination-based cloning using the multisite Gateway system (Hartley et al., 2000; Cheo et al., 2004) greatly simplifies the generation of expression constructs....
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...Gateway cloning technology is based on the att site-specific recombination system from lambda phage (Hartley et al., 2000)....
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