GENERAL SEARCHES
Our database can be searched in two ways:
- by GENE NAME: you may enter
either a systematic (e.g., YIL046W) or non-standard (e.g., MET30) gene name to
obtain a list of clones containing an mTn insertion affecting that gene.
- by CLONE ID: each strain in
our collection has been assigned a unique alphanumeric tag (e.g., V108A12). If
you already know the clone ID of a particular strain of interest, you may
enter that information here; otherwise, please search our database by gene
name.
Return to Main page
GENERAL SEARCH QUERY RESULTS
Search results are formatted as a table with
each row containing information for a given strain carrying the indicated
transposon insertion.
- Each strain is designated by its Clone ID : please click on this ID to receive a Composite Report of collected data for this clone —
including expression data, phenotypic data, and protein localization data.
- Insertion Point
identifies the site (codon
number) of transposon insertion within a given coding sequence.
- Total Protein Length indicates
mature protein length (number of amino acid) encoded by the
coding sequence into which the transposon has inserted.
Potential ORFs Disrupted by mTn
Insertion
A single transposon insertion event may disrupt any of a number of open reading frames (ORFs) within a given region of the yeast genome. For example, a nonannotated ORF (NORF) may overlap a previously annotated ORF; a transposon insertion within this overlapping region may, therefore, disrupt either the NORF or annotated ORF. In this case, both
the NORF and annotated ORF would be listed in our table.
- Insertion Point
identifies the site (codon
number) of transposon insertion within a given coding sequence.
- Total ORF Length
indicates the
mature protein length (number of amino acids) encoded by the coding sequence into
which the transposon has inserted.
- Repeated Gene:
an indication of whether the gene into which the transposon has inserted is
repeated within the yeast genome (in which case we would be unable to
conclusively identify its genomic site).
- LacZ Orientation: the lacZ gene (carried by
our mini-transposon) may be oriented either sense or antisense in respect to
its surrounding genomic context.
Insertion Point Data
A variety of underlying mTn insertion events are
responsible for b-gal activity within the strains constituting our
collection. We have classified these insertion events as follows:
- 3' region: transposon
insertion occurs within a region 200 bp 3' of the gene's translation
termination codon
- 5' region: transposon
insertion occurs within a region 200 bp immediately upstream of the gene's
initiator methionine codon
- Antisense: the inserted
transposon is oriented in the antisense direction within the gene
- Frame =+1: a productive
lacZ fusion has resulted from mTn insertion within the indicated coding
sequence in the +1 reading frame
- Frame = -1: a productive
lacZ fusion has resulted from mTn insertion within the indicated coding
sequence in the -1 reading frame
- In frame: a productive lacZ
fusion has resulted from an in-frame mTn insertion event within the indicated
ORF
- Non-protein-coding sequence
: we have observed some
productive lacZ fusions resulting from mTn insertion within genomic sequences
not expected to encode protein products (e.g., rDNA genes)
Gene Expression Data
To determine whether a given transposon-tagged gene is expressed during vegetative growth or sporulation, transposon-mutagenized yeast genomic DNA is transformed into the indicated background strain;
strains are then grown on rich medium and sporulation medium for approximately
17-20 hours. b-gal activity is
subsequently measured using a filter-based assay.
LacZ
expression levels are determined from the
observed intensity of blue staining after growth on medium containing X-gal.
Stain intensity is scored as follows:
- faint blue stain
- light blue color
- blue
- intense blue staining
Subcellular
Localization
Strains carrying a HAT-tagged protein were examined by indirect
immunofluorescence using monoclonal antibodies directed against the
transposon-encoded HA epitope.
- Resulting localization patterns have been
classified as follows:
- background: lack of
staining above a background level (some punctate background staining is
typically visible)
- buds: staining localized
to buds — sometimes observed specifically at the bud tip (see comments)
- cell neck: staining at
the mother-bud neck
- cell periphery: staining
localized to the outer cell membrane
- cyto.: whole-cell
staining
- cyto. dots: punctate
pattern of cytoplasmic staining (throughout the whole cell)
- cyto., non-nuclear:
cytoplasmic staining (excluding the nucleus)
- gran. cyto.: granular
(though not quite punctate) staining of the whole cell
- gran. cyto. non-nuclear:
granular (not punctate) staining of the cytoplasm — excluding the nucleus
- gran. nuclear: granular
(not punctate) staining of the nucleus
- mitochondria: visualized
as punctate cytoplasmic staining coincident with DAPI-staining
- nucleus: staining
localized to the nucleus
- nuclear rim / ER:
staining of the nuclear rim is often difficult to differentiate from
staining of the endoplasmic reticulum; therefore, we have grouped these
localizations together
- nucleolus: staining of
the nucleolus, visualized as a crescent-shaped structure occupying roughly
one-third of the nucleus
- spindle pole body:
visualized as a single dot in unbudded and budded cells; 2 dots (oriented
towards opposite ends) in a mitotic cell
- vacuolar rim: staining
localized to the periphery of vacuoles
- vacuole: staining coincident with a round,
non-nuclear area (appearing as a clear area when stained with
DAPI)
Fluorescent micrographs are available for any highlighted
localizations. Please click on the underlined localization entry to
view a corresponding IF image of yeast cells stained with the
DNA-binding dye DAPI (left-hand image) and the same cells stained by
indirect immunofluorescence using Anti-HA antibodies (right-hand image).
- Cell Stages and types are
classified as follows:
- all: localization not
restricted to any specific cell stage
- broken buds: staining
localized to a daughter bud separated from its mother cell
- budded: staining of cells
undergoing budding; immunofluorescence not localized to either the mother
cell or daughter bud
- buds: staining localized
to a bud still attached to its mother cell (no staining of the mother)
- budded-mother:
immunofluorescence localized to a mother cell undergoing budding (no
staining of the daughter bud)
- mitosis: staining
localized to cells undergoing mitosis (not restricted to either mother or
daughter cells)
- mitosis-daughter:
staining of mitotic cells; restricted to daughters
- mitosis-mother: staining
of mitotic cells; restricted to mother cells
- unbudded: localization to unbudded
cells
Please bear with our data — we endeavor to classify observed staining
patterns as thoroughly as possible
- % of
cells staining: an indication of the fraction of cells exhibiting the
observed staining pattern
- Stain
Intensity: the observed intensity of immunofluorescence is estimated
as follows:
- 1 = weak staining (comparable to background
levels)
- 2 = definite staining above background
- 3 = strong staining
- 4 = very intense staining
- Trials: the number of times a given
strain has been examined by immunofluorescence with anti-HA antibodies; all
strains exhibiting discrete cellular or cytoplasmic localizations were tested
at least twice.
Disruption Phenotype Data
Haploid strains (each carrying an independent mTn insertion) were screened for disruption phenotypes using the following assays (growth conditions):
- HapTra: cell inviability of
haploid transformants
- HHIG: hyperhaploid invasive
growh mutants
- White: white/red colony
color on YPD (indicating genes potentially functioning in cell respiration)
- YPD37C: YPD at 37°C
(indicating temperature-sensitive mutants)
- YPD11C: YPD at 11°C
(indicating temperature-sensitive mutants)
- 8Caff: YPD + 8 mM caffeine
(indicating genes potentially functioning in cell wall maintenance/biogenesis)
- 10Ben: Benomyl
hypersensitivity as assayed by growth on YPD + 10 mg/ml benomyl
(indicating genes potentially functioning in microtubule dynamics)
- 20Ben: Benomyl resistance
as assayed by growth on YPD + 20
mg/ml benomyl
- MethBl: YPD + 0.001%
methylene blue (indicating genes potentially functioning in cell wall
maintenance/biogenesis)
- BCIP: YPD + BCIP
(indicating genes potentially functioning in cell wall maintenance/biogenesis)
- 01MMS: YPD + 0.008% MMS
(indicating genes potentially functioning in processes of DNA metabolism)
- 2EGTA: YPD + 2 mM EGTA
(indicating cation-sensitive mutants)
- 12Calc: calcofluor
hypersensitivity as assayed by growth on YPD + 12
mg/ml
calcofluor (indicating genes potentially functioning in cell wall
maintenance/biogenesis)
- 67Calc: calcofluor
resistance as assayed by growth on YPD + 66.7
mg/ml
calcofluor (as above, indicating genes potentially functioning in cell wall
maintenance/biogenesis)
- 75mMHU: YPD + 75 mM
hydroxyurea (indicating genes potentially functioning in processes of DNA
metabolism)
- CyhS: cycloheximide
hypersensitivity as assayed by growth on YPD + 0.08
mg/ml
cycloheximide
- CyhR: cycloheximide
resistance as assayed by growth on YPD + 0.3
mg/ml
cycloheximide
- 46HygR: YPD + 46
mg/ml hygromycin (indicating genes potentially functioning
in cell wall maintenance/biogenesis)
- 003SDS: YPD + 0.003% SDS
(indicating genes potentially functioning in cell wall maintenance/biogenesis)
- 9NaCl: YPD + 0.9 M NaCl
(identifying salt-sensitive mutants)
- YPGly: assay for growth
on YPGlycerol (containing glycerol as the sole carbon source) in order to
identify genes potentially functioning
in cellular respiration.
- Score: growth of each transformant in a selective
plate is compared with growth of the same transformant in a YPD control plate
during a period of 2-6 days. Differences in growth are scored as follows:
- ND = no difference as
compared to growth on YPD
- weak = weak difference in
growth
- medium =
medium
difference in growth
- strong =
strong
difference in growth
- wt and not wt
refer to "wild-type" and "not wild-type" growth patterns
respectively
Background strain and its ploidy are also included per entry:
- 1n = haploid
- 2n = diploid
Search NORF Data
In annotating the S. cerevisiae genome, an arbitrary
lower size limit of 100 amino acids was adopted as convention in the absence of
specific information to the contrary. Therefore, typically, open reading frames
<100 codons in length are not annotated. These non-annotated ORFs (NORFs),
however, are capable of being translated; some may be of significant biological
relevance. Our transposon-tagging approach is an effective means of identifying
NORFs -- we have collected data concerning these NORFs for easy access using
this search form.
NORFs are sometimes found overlapping (or within) larger
annotated ORFs; to specifically search for a NORF located within (or in the
vicinity of) a given gene, please enter that gene name within the "genome
region" field.
For an explanation of indicated categories, please
read our insertion point data help
text.