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General Information
Symbol
PBac{UAS-DF-Syt1}
FlyBase ID
FBtp0134946
Feature type
transgenic_transposon
Component Allele(s)
Size
Expression Data
Associated insertion(s)
Molecular map
Description and Uses
Location-dependent role
Description
CV term
Qualifiers and info
Reference
transposon
Tni\piggyBac
Construct components
Product class / Tool use(s)
Description

UASt regulatory sequences drive expression of a sensor that is composed of Syt1 fused at its N-terminal end to a mCherry-pHluorinSE fusion. The presence of Syt1 targets the protein to the synaptic vesicle membrane (the dual fluorophore faces the vesicle lumen). The mCherry-pHluorinSE dual fluorophore can act as an acidification sensor in vivo as pHluorinSE is pH sensitive but mCherry is pH-resistant; red fluorescence indicates an acidic environment (pH < 6), while yellow fluorescence indicates a pH greater or equal to 7. In addition, the C-terminal pHluorinSE moiety is expected to be more exposed to proteases than mCherry, such that the dual fluorophore can also be used as a degradation sensor by observing fluorescence in fixed samples; yellow fluorescence indicates both fluorophores are intact, red fluorescence indicates degradation of the C-terminal pHluorinSE moiety.

Sequence Data
Sequence (FB)
Associated Sequence Data
DNA Sequence
Extent
Segments and Size
Total Size
Left end
Right end
 
Segments
Num
Orientation
Symbol
Reference
Features
CV term
Qualifiers and info
Reference
Component Alleles
Reference(s)
Molecular data

UASt regulatory sequences drive expression of a sensor that is composed of Syt1 fused at its N-terminal end to a mCherry-pHluorinSE fusion. The presence of Syt1 targets the protein to the synaptic vesicle membrane (the dual fluorophore faces the vesicle lumen). The mCherry-pHluorinSE dual fluorophore can act as an acidification sensor in vivo as pHluorinSE is pH sensitive but mCherry is pH-resistant; red fluorescence indicates an acidic environment (pH < 6), while yellow fluorescence indicates a pH greater or equal to 7. In addition, the C-terminal pHluorinSE moiety is expected to be more exposed to proteases than mCherry, such that the dual fluorophore can also be used as a degradation sensor by observing fluorescence in fixed samples; yellow fluorescence indicates both fluorophores are intact, red fluorescence indicates degradation of the C-terminal pHluorinSE moiety.

Expression Data
Reporter Expression
Additional Information
Statement
Reference
 
Marker for
Reflects expression of
Reporter construct used in assay
Progenitors and Descendants
progenitor(s)
descendant(s)
Comments
Stocks (0)
External Crossreferences and Linkouts ( 0 )
Synonyms and Secondary IDs (3)
Reported As
Symbol Synonym
PBac{UAS-DF-Syt1}
UAS-DF-Syt1
UAS-mCherry-pHluorin-Syt1
Secondary FlyBase IDs
    References (2)