a+ --> a- mutation (forward mutation)

1. a+ --> a- mutation (forward mutation) a- --> a+ reverse mutation (reversion)

2. Presence of pink pigment + / - Incomplete dominancethe term used to describe the general casein which the phenotype of a heterozygote is intermediate between those of the two homozygotes,on some quantitative scale of measurement Four-o’clock plants

3. Point mutations at the molecular level Purine replaced by a different purine; pyrimidine replaced by a different pyrimidine: TRANSITIONS Purine replaced by a pyrimidine; pyrimidine replaced by a purin: TRANSVERSIONS

6. Animation ed.9: 9.2&9.17 TRANSLATION

8. Animation ed.9: 9.2&9.17 TRANSLATION

13. Now, insertions and deletions of base pairs:

15. Selection of auxotrophs by filter enrichment Forward mutation-A mutation that converts a wild-type allele into a mutant allele

16. Reverse mutation-The production of a wild-type gene from a mutant gene

18. Intragenic suppressor Equivalent reversion UCC (Ser)forwardUGC (Cys)reverseAGC (Ser) Wild type Mutant Wild type CGC (Arg, basic) forwardCCC (Proline) reverseCAC (His, basic) Wild type Mutant Pseudo-wild type

19. Intragenic suppressor

20. Intergenic suppressor

21. Intergenic suppressor Nonsense suppressor

23. non-essential Chromosome Fragment SUP11 M WT ade2-101 CIN mutant ade2-101 ade1-101 Chromosome transmission fidelity (Ctf) assay ade2-101 kar3D sic1D rad50D xrs2D

24. Animation ed9: 9.19a Nonsense mutation

25. Animation ed9: 9.19b Nonsense suppressor

26. Animation 9.19c Nonsense suppression rodns and suppressor-tRNA together give WT phenotype

27. Coding Regulatory

30. Using genomic sequence to find a specific gene

31. When doing GENETIC mapping,Molecular Markers can be used as a locus Single Nucleotide Polymorphisms (SNPs) AACGTCATCG vs. AACGTTATCG Microsatellites (variable # of short repeats) CGCGCG vs. CGCGCGCGCG vs. CGCG Restriction Fragment Length Polymorphism (RFLP) SNP leading to a loss/gain of a restriction cut site

32. When doing GENETIC mapping,Molecular Markers can be used as a locus Almost all SNPs, Microsatellites, etc. are SILENT, and there are millions of them They are mile-markers, not destinations! אבני דרך, ולא יעדים!

33. A specific gene, the breast cancer gene BRCA1 was found By using the genomic map at increasing levels of resolution

34. Is there linkage between a mutant gene/phenotype and a SNP? USE standard genetic mapping technique, with SNP alternative sequences as “phenotype” B= bad hair, Dominant B 2’ / b 2 SNP1 ..ACGTC.. SNP1’ ..ACGCC.. SNP2 ..GCTAA.. SNP2’ ..GCAAA.. SNP3 ..GTAAC.. SNP3’ ..GTCAC.. X 1/1’1/1 B/b b/b 2/2’ 2/2 3/3’ 3/3 2/2’ 47% 2/2 3% 2/2’ 3% 2/2 47% 3/3’ 25% 3/3 25% 3/3’ 25% 3/3 25% 1/1’ 25% 1/1 25% 1/1’ 25% 1/1 25% B/b B/b b/b b/b SO…B is 6 cM from SNP2, and is unlinked to SNP 1 or 3

35. Is there linkage between a mutant gene/phenotype and a SNP? USE standard genetic mapping technique, with SNP alternative sequences as “phenotype” B= bad hair, Dominant SNP1 ..ACGTC.. SNP1’ ..ACGCC.. SNP2 ..GCTAA.. SNP2’ ..GCAAA.. SNP3 ..GTAAC.. SNP3’ ..GTCAC.. X 1/1’1/1 B/b b/b 2/2’ 2/2 3/3’ 3/3 We have the ENTIRE genome sequence of mouse, so we know where the SNPs are Now-do this while checking the sequence of THOUSANDS of SNPs SO…B is 6 cM from SNP2, and is unlinked to SNP 1 or 3

36. The logic of creating sequence map of the genome

37. Physical maps are maps of the order, overlap, and orientation of physically isolated pieces of the genome-in other words, maps of the distribution of the cloned genomic DNA from genomic clone libraries

38. Part of the automated production line of a major human genome sequencing center

39. A specific gene can be found in the genomic sequence by matching linkage and cytological maps with the Genome sequence

44. CGCGCG vs. CGCGCGCGCG vs. CGCG

48. Complementation groups Mutagenesis Plate to select for phenotype of interest

49. Complementation groups First, we need to catalogue our mutants to complementation groups (Total of 138 mutants were isolated in the original CTF screen). Mutant#1 Mutant#2 Diploid x x x GENE1 x GENE1 GENE1 Mate GENE1 Diploid still shows CTF phenotype Mutant#1 and Mutant#2 are mutated in the same gene Same complementation group

50. Complementation groups Mutant#3 Mutant#4 Diploid x GENE9 x x GENE9 x GENE10 GENE9 Mate GENE10 GENE9 GENE10 GENE10 Diploid dont show CTF phenotype Mutant#3 and Mutant#4 are mutated in different genes Different complementation groups