This Week

• Continue Arabidopsis Development Screen,

– discussion,

– assignment (20 pts., submit one per group),• describe your putants, describe and illustrate the growth and

development, through week 10 (due December 7th),– submit seeds in a sealed envelope (i.e., pick a fertile putant),

• Continue HFR experiment.

ScreensMulti-Quarter Project

Conditional Screen

• Rescue putants,• Complementation,• If known, submit to

Arabidopsis Biological Resource Center.

• If novel, map, then submit.

Developmental Screen

• Collect seeds,• Retest progeny,

– statistical relevance,

• Cross with wt,– F1, F2 segregation?

• Map,• Clone?

Classical Mapping

Cross an organism with a trait of interest to homozygous mutants of known mapped genes.

Then, determine if segregation is random in the F2 generation,

• if not, then your gene is linked (close) to the known mapped gene.

target

What recombination frequency do you expect between the target and HY2?

What recombination frequency do you expect between the target and TT2?

Molecular Markers

ScreensMulti-Quarter Project

Conditional Screen

• Rescue putants,

• Complementation,

• If known, submit to Arabidopsis Biological Resource Center.

• If novel, map, then submit.

Developmental Screen

• Collect seeds,• Retest progeny,

– statistical relevance,

• Cross with wt,– F1, F2 segregation?

• Map,• Clone?

Assignment(due December 7th)

– assignment (20 pts., submit one per group),

• describe your putants, describe and illustrate the growth and development, through week 10,

– dates, description, drawings, etc.,– special conditions, etc.

• submit seeds in a sealed envelope (i.e., pick fertile putants),

– staple envelopes to your report

lac (7)

car::tn10 (0.6)tetR

car (0.6)tetS

rpoB- (90)rifR

rpoB+ (90)rifS

metB- (89)

metB+ (89)

cysG- (72) cysG+ (72)

gyrA- (48)

nalS

gyrA+ (48)

nalR

1st Selection: tetR, nalR

Plate A proline, arginine, uracil

Plate B proline, arginine, uracil,methionine

Plate C proline, arginine, uracil,cysteine

Plate D proline, arginine, uracil,methionine, cysteine

Supplements Marker Selected

met, cys

cys

met

no selection

Selection Scheme

lac (7)

car::tn10 (0.6)tetR

car (0.6)tetS

rpoB- (90)rifR

rpoB+ (90)rifS

metB- (89)

metB+ (89)

cysG- (72) cysG+ (72)

gyrA- (48)

nalS

gyrA+ (48)

nalR

Met +, Cys + Met +, Cys - Met -, Cys +Met -, Cys -

Met+ Cys+ Plates: A __ B __ C __ D __

lac (7)

car::tn10 (0.6)tetR

car (0.6)tetS

rpoB- (90)rifR

rpoB+ (90)rifS

metB- (89)

metB+ (89)

cysG- (72) cysG+ (72)

gyrA- (48)

nalS

gyrA+ (48)

nalR

Met+ Cys- Plates: A __ B __ C __ D __Met- Cys+ Plates: A __ B __ C __ D __Met- Cys - Plates: A __ B __ C __ D __

lac (7)

car::tn10 (0.6)tetR

car (0.6)tetS

rpoB- (90)rifR

rpoB+ (90)rifS

metB- (89)

metB+ (89)

cysG- (72) cysG+ (72)

gyrA- (48)

nalS

gyrA+ (48)

nalR

Met_, Cys+ Plates: A __ B __ C __ D __

lac (7)

car::tn10 (0.6)tetR

car (0.6)tetS

rpoB- (90)rifR

rpoB+ (90)rifS

metB- (89)

metB+ (89)

cysG- (72) cysG+ (72)

gyrA- (48)

nalS

gyrA+ (48)

nalR

Met_, Cys- Plates: A __ B __ C __ D __

lac (7)

car::tn10 (0.6)tetR

car (0.6)tetS

rpoB- (90)rifR

rpoB+ (90)rifS

metB- (89)

metB+ (89)

cysG- (72) cysG+ (72)

gyrA- (48)

nalS

gyrA+ (48)

nalR

Met+, Cys- Plates: A __ B __ C __ D __