Genetics 2165 Tutorial 5

1. a) Eight independent tyr- mutants are isolated. Complementation tests are performed in all pairwise com inations !" # complementation$ - # no complementation). 1 1 2 % & 5 6 ' ( 2 " % " " & " 5 " " " 6 " " " " ' " " " " ( " " " " -

) )ow would you determine colonies that are tyr-* !2m+s)

Plate the bacteria on a medium that contains tyrosine, replica plate on a medium lacking tyrosine. Colonies that do not grow in plate two are tyr-.

c) ,ith the aid of a diagram- suggest the num ers of genes present- indicating which mutations are present on each genes. !&m+s)
3 genes: 1,4,8 in gene-1 , ! in gene", # in gene-3 $ene 3 is a deletion mutatant spanning part o% gene one and all o% gene 3.

&

'

.r % 2 5 1 & ( 6 '

2. a) / cross is made etween an )fr strain of E. coli that is met" thi" pur" and an 0- strain that is met- thi- pur-. 1nterrupted mating studies show that thi" enters the recipient last- so that thi" recom inants in the 0- ac+ground are selected on a medium containing supplements that satisfy only the purine and methionine re2uirements. These recom inants are tested for the presence of met" and pur" alleles. The following num ers of indi3iduals are found for each genotype.

thi" met" pur" %24 thi" met" purthi" met- pur" thi" met- pur6 5& (4

b& ,hy was thiamine left out of the medium* !%m+s) 'ack o% thiamine selects bacterial colonies that ha(e the gene to synthesi)e thiamine. *ince interrupted mating e+periments indicate that thiamine enters last, selection o% thiamine positi(e genotypes would ensure that all earlier genes ,genes more pro+imal to the origin than thiamine& would ha(e had an e-ual chance o% integrating into the genome. .his would allow one to use recombination mapping in bacteria.

c& ,hat is the gene order* !&m+s)

.he central gene is the gene that is unrecombined in the genotype with the lowest %re-uency. .he genotype with the lowest %re-uency is met/ thi/ pur-. .he unrecombined gene in this genotype is pur-. 0ence pur must be the central gene. *ince thi/ enters last. .he gene order must be met-pur-thi.

d& ,hat are the map distances in recom ination units

etween the genes* 5raw illustrations to indicate the 3arious methods of recom ination. !' m+s6% m+s for diagram 1 mar+ each- 2m+s for each calculation- 2m+s for the map)

1rom the diagrams a& 2et/ pur/ thi/ 2et- pur- thior b& .hi/ pur/ met/ .hi- pur- met*C345

a& 2et/ pur/ thi/ 2et- pur- thi-

or

.hi/ pur/ met/ .hi- pur- met-

*C3455

4earrange the genes since the order is known: 6& 2et- pur- thi/ 7 88 2et- pur / thi/ 7 !4 2et/ pur- thi/ 7 6 2et/ pur/ thi/ 7 %24 4C : ,45& 7 ;sco ,45& / dco < total= + 188 7 ;!4/ "< 4"8= + 188 7 13.84 : 4C: ,455& 7 ;88 / "<4"8= + 188 7 18.#: $ene map met pur thi sco 455 sco 45 dco ,45 9 455&

13.84 mu

18.# mu

.r 7)
2et- pur- thi/ 7 88 2et- pur / thi/ 7 !4 2et/ pur- thi/ 7 6 2et/ pur/ thi/ 7 %24 sco 45 sco 455 dco ,45 9 455&

4C : ,45& 7 ;sco ,45& / dco < total= + 188 7 ;88/ "< 4"8= + 188 7 18.# : 4C: ,455& 7 ;!4 / "<4"8= + 188 7 13.84:

thi 18.# mu

pur 13.84 mu

met

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