Beruflich Dokumente
Kultur Dokumente
Yongheng Liang
College of Life Science
Nanjing Agricultural University
Nanjing 210095
Tel/Fax: 025-8439 6376
Homepage: http://lfc.njau.edu.cn/common.asp?d=25&m=3&i=47
Yeast Biology
Whats yeast?
Saccharomyces cerevisiae
(budding yeast/
Bakers yeast )
Schizosaccharomyces pombe
(fission yeast)
Yeast Basics
Part II.
Part III.
Part IV.
Part V.
Part VI.
Part I.
Yeast Basics
Mitochondria
Vacuole
Nucleus
Cell wall
Vesicle
Eduard Buchner
Leland H. Hartwell
WT, 37C
sec1-1, 37C
1h
sec1-1, 24C
sec1-1, 37C
2h
Novick P, Schekman R: Secretion and cell-surface growth are blocked
in a temperature-sensitive mutant of Saccharomyces cerevisiae. Proc
Natl Acad Sci USA 1979; 76:1858-1862.
Yeast colony
Diploid cell
Volume (m3)
70
120
Diameter (m)
5-6
FM4-64
Merged
Budding video
Is asexual reproduction
a mitosis or an amitosis?
Part II.
FIVE IS
2. Incubation
1. Inoculation
4. Inspection
5. Identification
3. Isolation
Streak
Dilute and
pour plate
Incubator
flask rack
rotor drum
Channel pipette
200
180+20=200
1/100
180+20=200
1/1000
180+20=200
1/10000
180+20=200
96-well plates
PhC
DIC
Other microscopies
Confocal
microscopy
Elo3-GFP
endoplasmic reticulum.
Nile red staining
of lipid droplets in obese yeast
Tansmission
electron-microscopy
Leica Ultracut
JEM-1011
Scanning electron-microscopy
JEOL SEM
SEM for
budding yeast
Dissection microscopy
Part III.
Macromolecular
constituents of
yeast comprise
proteins,
glycoproteins,
polysaccharides,
polyphosphates,
lipids, and
nucleic acids
DIC
DAPI staining
ypt1ts
DIC
Live microscopy
DAPI
ypt1ts
Htb1-CFP
-Cell wall
PeriplasmPMNucleus
Vacuole
ER-
Autophsgic
bodies
Plasma membrane
Nucleus
Vacuole
ER
DIC
GFP-Pmp2
DIC
WT
mutant
GFPSnc1
Sec7DsRed
merged
Ypt7
Ypt51,52,53
Ypt11
Ypt6
Ypt1
ER
E
E
Ypt31,32
Sec4
secretory PM
vesicles
GOLGI
Endocytosis
Vacuoles
DIC
GFP
FM4-64
merged
WT
mutant
Category of Autophagy
Mictophagy (Mitochondria)
Chlorophagy (Chlorophyll)
Pexophagy (Peroxisome)
merged
GFP
FM4
DIC
WT
ypt51
ypt52
ypt53
ypt51GFP-Atg8
SD-N
WT
ypt51
DIC
WT
ypt51
ypt51
ypt52
Atg8
Ape1
merged
A model of cytoskeleton
Part IV.
Budding
How to bud?
Mutant phenotypes in
budding
Wild-type haploid
an axial budding pattern
Wild-type diploid
ste20-delta diploid
bud7-delta diploid
Spindle dynamics
Septins
START
Identification of cdc28
35S-methionine
Leland H. Hartwell
a
a
Meiosis
uThe life cycle of S. cerevisiae
normally alternates between
diplophase and haplophase. Both
ploidies can exist as stable cultures.
uIn heterothallic strains, haploid
cells are of two mating types, a and
. Mating of a and cells results in
a/ diploids that can undergo
meiosis.
uThe four haploid products
resulting from meiosis of a diploid
cell are contained within the wall of
the mother cell (the ascus).
Digestion of the ascus and
separation of the spores by
micromanipulation yield the four
haploid meiotic products.
ascus
2N
Part V.
Vp
s8
Vp
s9
Yp
1
5
t
8
s
p
9
s
p
Vps34
-Leu-Trp
-Leu-Trp-His
Yp
1
5
t
Yeast Basics
Part II.
Part III.
Part IV.
Part V.
Part VI.
Eduard Buchner
Part VI.
Yeast
H2O
H2O
Sugar
H2O
Yeast 0.5g
Sugar 1g
H2O 5ml
Before
After
WT
ypt51
Atg21-GFP
RFP-Atg8
merged
2:00-3:30
3:30-5:00
Room
Biology (20)
Chemistry (20)
C5023
for yeast
balloon exp
C6012
C6012
for fluorescence
observation
C5023
Ypt
ole
Vacu
Thank
You!