GFP expression in Drosophila tissues: time requirements for formation of a fluorescent product.
We analyzed the acquisition of fluorescence of a GFP fusion protein expressed from gene constructs designed to regulate either the time of translation of the protein or the time of gene transcription. Both gene constructs expressed a fusion protein with wild-type GFP appended to the N-terminus of the Bicoid (Bcd) transcription factor. One gene construct, P[gfp-bcd] expressed an mRNA that was initially translated at the time of egg deposition. GFP-Bcd fluorescence could first be detected in anterior nuclei in the embryo at syncytial blastoderm stage, within 1.5-2.5 h after translation commenced. The second gene construct, P[hs-gfp-bcd], utilized the hsp70 promoter to regulate the induction of gene expression. In larval salivary glands, GFP-Bcd fluorescence could first be detected in nuclei after a 40-min recovery period following a 10-min heat shock induction of gene expression. These estimates of the time required for the acquisition of a fluorescent GFP product in Drosophila tissues are less than suggested by previously published reports.
Hazelrigg
T
T
Department of Biological Sciences, Columbia University, New York, New York 10027, USA.
Liu
N
N
Hong
Y
Y
Wang
S
S
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004330:Drosophila / Q000196:embryology / Q000235:genetics* / Q000378:metabolism; D029721:Drosophila Proteins; D018507:Gene Expression Regulation, Developmental; D017344:Genes, Insect; D049452:Green Fluorescent Proteins; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics; D018398:Homeodomain Proteins / Q000096:biosynthesis / Q000235:genetics; D007700:Kinetics; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics*; D008856:Microscopy, Fluorescence; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D015534:Trans-Activators / Q000096:biosynthesis / Q000235:genetics
D017931:DNA Primers; D029721:Drosophila Proteins; D018840:HSP70 Heat-Shock Proteins; D018398:Homeodomain Proteins; D008164:Luminescent Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; C056478:bcd protein, Drosophila; D049452:Green Fluorescent Proteins