Background and notes for yeast genetics





Nomenclature

Yeast gene names are three letters followed by a number and are written in italics.
  i.e.     URA3     (the third gene identified that is required for uracil biosynthesis)
  CDC44    (the 44th cell division cycle mutant that was identified)

The one exception to this nomenclature is the yeast mating type locus which is designated;
 Mat a      : haploid a type cells
 Mat a     : haploid a type cells
 Mat a/ a : diploid cells

Normal, unmutated, functional alleles of genes are designated as wild type, and can have the + superscript.  i.e.  CDC44 +

If a gene is not listed in the genotype if a strain, it is assumed to be wild type.

Mutant genes are followed by an allele number that specifies the mutation.
 i.e.   cdc44-1  (first mutant allele, happens to be a point mutation at amino acid 513)
         lys2-*301  (encodes for a Lys2 protein containing a 301 amino acid deletion)

Capital letters signify dominant genes, lower case letters for recessive genes
 i.e.    LEU2      dominant wild type allele
  SUP4-1   dominant mutant allele
  cdc44-1   recessive mutant allele

If a chromosomal copy of a gene has been deleted and replaced with another gene, it is indicated with two colons (::) as follows;
      CDC44::LEU2  (CDC44 gene is absent and LEU2 gene is located where CDC44 was)

However, if a gene is placed next to another gene on a chromosome without destroying its function, it is indicated as follows;
 cdc44-1.URA3  (URA3 gene has been integrated next to cdc44-1  gene without  destroying cdc44-1 or URA3 function)

Plasmids genotypes are written in parantheses   (URA3  CDC44)

For diploid cells, both copies of the genes are indicated as follows;

 Mat a/ a        CDC44   ura3-52    leu2-1
             cdc44-1    ura3-52    LEU2

Proteins are written in normal type, with the first letter capitolized and have the letter p following gene name,
 i.e.   Cdc44p     =  protein encoded by wild type CDC44 gene
  Cdc44-1p  = protein encoded by mutant cdc44-1 gene

Phenotypes are written in normal type, and can have r/s +/- designations
 i.e. Ura-  =  strain cannot grow in absence of uracil  (Ura auxotroph)
  ts-     =  strain cannot grow at elevated temperatures  (35°C)
           Mmss  = strain is sensitive to MMS

Plates

YPD = rich media made from yeast extract (supports growth of all strains)
SC  = synthetic complete media (synthetic media with all nutrients and amino acids etc)
SD  = synthetic deficient media  (has only salts and sugars)
SC - His = synthetic media with all the nutrients and amino acids to grow except histidine

For our purposes,   SC =  SD +  His,  Lys,  Leu,  Ura,  Ade,  Tyr,  Trp

i.e. therefore  SC - Ura  =  SD + His,  Lys,  Leu,  Ade,  Tyr,  Trp

 

I. Segregation of 2 unlinked genes

  Genotype    Phenotype  Class

  Mat a     cdc44-1     LEU2  cs-,  Leu+  Parental
crossed with
  Mat a    CDC44      leu2-1  cs+,   Leu-  Parental

gives  diploid Mat a/ a      cdc44-1      LEU2     cs+, Leu+
                 CDC44      leu2-1

upon sporulation can give haploids like (remember meiosis is one round of DNA replication followed by two rounds of chromosome segregation).  Each spore gets one copy of each gene.
 
Tetrads Genotype  Phenotype     Class   Type

1  cdc44-1  LEU2 cs-,  Leu+        Parental Ditype
  cdc44-1  LEU2 cs-,  Leu+     “
  CDC44  leu2-1 cs+,  Leu-  Parental
  CDC44  leu2-1 cs+,  Leu-     “

2  cdc44-1  leu2-1 cs-,  Leu-  Non-Parental Ditype
  cdc44-1  leu2-1 cs-,  Leu-     “
  CDC44  LEU2  cs+, Leu+  Non-Parental
  CDC44  LEU2  cs+, Leu+     “

3  cdc44-1  LEU2 cs-,  Leu+  Parental  Tetratype
  cdc44-1  leu2-1 cs-,  Leu-  Non-Parental
  CDC44  LEU2  cs+,  Leu+  Non-Parental
  CDC44  leu2-1 cs+,  Leu-  Parental

4  cdc44-1  leu2-1 cs-,  Leu-  Non-Parental Tetratype
  cdc44-1  LEU2 cs-,  Leu+  Parental
  CDC44 leu2-1  cs+,  Leu-  Parental
  CDC44 LEU2  cs+,  Leu+  Non-Parental

5  cdc44-1  LEU2 cs-,  Leu+  Parental Tetratype
  cdc44-1 leu2-1  cs-,  Leu-  Non-Parental
  CDC44  leu2-1 cs+,  Leu-  Parental
  CDC44 LEU2  cs+,  Leu+  Non-Parental

6  cdc44-1 leu2-1  cs-,  Leu-  Non-Parental Tetratype
  cdc44-1  LEU2 cs-,  Leu+  Parental
  CDC44 LEU2  cs+,  Leu+  Non-parental
  CDC44  leu2-1 cs+,  Leu-  Parental

 In all, ratio is:  1 Parental Ditype:  1 Non-parental Ditype: 4 Tetratype
 
 If genes are unlinked, then each one of these cases is equally likely (alleles segregate randomly).  Overall, each class of phenotype (i.e. cs-,  Leu+) accounts for 6/24 or 25% of progeny as would be expected (0.5 x 0.5).
 

II.  Segregation of two linked genes

Tetrads Genotype  Phenotype     Class   Type

1  cdc44-1  LEU2 cs-,  Leu+        Parental Ditype
  cdc44-1  LEU2 cs-,  Leu+     “
  CDC44  leu2-1 cs+,  Leu-  Parental
  CDC44  leu2-1 cs+,  Leu-     “
 

 If the CDC44 and LEU2 genes are tightly linked to one another (physically very close to each other on the chromosome) then the resulting phenotypes of the spores would be the same as in the parental strains.  Only a recombination event between the CDC44 and LEU2 genes could give rise to a different phenotype of spore (i.e. cs+,  Leu+).  Therefore, the further away the genes are, the more likely a recombination event, the more tetratype products of meiosis.

    In all ratio is:  many Parental Ditypes: very few Non-parental ditypes:  few tetratypes