Kit 11

Streptomycin Point Mutants in S. typhimurium

One method to locate a gene on the chromosome if it has a selectable phenotype is to transfer it by conjugation and look for linkage to known genes. The strain with the selectable marker e.g., Tn10 with Tet-res can be made Flac(+) by conjugation and selection for Lac(+); this strain can be mated as donor to a strain with a known auxotrophic mutation (e.g., trp) with selection for Tet-res (from the donor chromosome) and Str-res (counter-selection against the donor cell), and the linkage between Tet-res and the Trp(- )phenotype can be determined. A set of about 40 Str-res strains, each with a single auxotrophic mutation at locations around the chromosome, has been constructed, as listed below.

 Min

 Strain #

 Gene

 Auxotrophy
 0  SA3661  thrA  threonine
 2  SA3646  pyrA  uracil and arginine
 3  SA3662  leuB  leucine
 5  SA3630  panB  pantothenic acid
 7  SA3631  proB  proline
 8.5  SA3632  proC  proline
 12  SA3663  purE  adenine
 14  SA320 lip   lipoic acid
 18.5  SA3633  bio  biotin
 21  SA3655  pyrD  uracil
 23  SA3658  pyrC  uracil
 25  SA3634  purB  adenine
 28  SA3635  aroD  tyrosine, tryptophan, phenylalanine
 33.5  SA3636  pyrF  uracil
 34  SA3637  trpC tryptophan
 42  SA3647  hisH histidine
 44  SA3648  metG  methionine
 47  SA3638  purF  adenine
 50  SA3649  cysA  cysteine
 51  SA3656  purC adenine
 54  SA3639  glyA  glycine
 55  SA3657  tyrA  tyrosine
 60  SA3650  cysC  cysteine
 61  SA3640  argA  arginine
 68  SA3651  argG  arginine
 71  SA3667  aroE  tyrosine, tryptophan, phenylalanine
 72  SA3664  argD  arginine, proline
 73  SA3668  aroB  tyrosine, tryptophan, phenylalanine
 79  SA3641  cysE  cysteine
 79.5  SA3840  pyrE  uracil
 83  SA3653  ilvE  isoleucine, valine
 84  SA3645  metE methionine
 87.5  SA3642  metB  methionine
 88  SA3654  argE  arginine
 89  SA3665  purD adenine
 96  SA3666  purA  adenine
 97  SA3660  argI  arginine
 98  SA3643  pyrB  uracil
 99.5  SA3644  serB  serine

 

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