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Mutation detection systems navi
- 1. PREPARED BY NAVEENA GIRISH DEPARTMENT OF PLANT SCIENCE CENTRAL UNIVERSITY OF KERALA
- 2. 2 TYPES VISIBLE SLC BIOCHEMICAL SLC WIDELY USED Uses 5 or 7 loci Uses up to 20 enzymes
- 3. T O DETECT QUANTITATIVE MUTATION IN MAMMALIAN GERMLINE CELLS A visible specific locus mutation is a genetic change that alters factors responsible for coat color and other visible characteristics of certain mouse strains principle to cross individuals who differ with respect to the genes present at certain specific loci, so that a genetic alteration involving the standard gene at any one of these loci will produce an offspring detectably different from the standard heterozygote. FEMALE HOMOZY GOUS MALE + TEST REAGENT PROGENY SCREENING FOR MUTATION
- 4. BIOCHEMICAL SPECIFIC LOCUS TEST A biochemical specific locus mutation is a genetic change resulting from a DNA lesion causing alterations in proteins that can be detected by electrophoresis methods. The principle of the MBSL is that heritable damage to the genome can be detected by electrophoresis analysis of proteins in the tissues of the progeny of mice treated with germ cell mutagens Treated males are then mated to untreated females to produce F1 progeny. Both blood and kidney samples are taken from progeny for electrophoresis analysis. Up to 33 loci can be examined by starch-gel electrophoresis and broad-range isoelectric focusing. Mutants are identified by variations from the normal electrophoresis pattern. Presumed mutants are bred to confirm the genetic nature of the change.
- 5. Single generation & visual detection is used to identify mutation – SO USEFUL OFFSPRING EXAMINATION BIRTH AND WEANING TISSUE SAMPLING ELECTROPHORESIS MUTANT IDENTIFICATION BLOOD -6 LOCI KIDNEY – 27 LOCI
- 6. AMES TEST Industrial chemicals potentially harmful to humans We can know it using lab animals But it is time consuming and expensive Bruce Ames 1974 PRINCIPLE Damage to DNA mutation Cancer 90% carcinogen = mutagen Mutation in bacteria = carcinogenesis in humans
- 7. I used Salmonella typhimurium with dna repair system inactivated and defects in lipopolysacharide 4 strains used
- 8. carcinogenic compounds in hair dyes removed by the help of Ames test
- 9. One of the four strains detect base pair substitutions Other three detect frameshift mutation
- 10. DROSOPHILA MELANOGASTER DEFINED MUTATION RATE INTIAL EXPERIMENTS WAS FAIL BUT 1927 BERLIN CONFERENCE HE SURPRISED OTHERS X- RAYS INDUCES MUTATION 7 YEARS BUT HE SAID NO QUALITATIVE DIFFERENCE BETWEEN SPONDANEOUS AND INDUCED MUTATIONS
- 11. ClB METHOD – TO STUDY MUTATIONS HAPPENING ON X CHROMOSOME
- 12. He received Nobel prize in 1945
- 13. MULERS -5 METHOD
- 14. TO TEST X CHROMOSOME The progeny of F2 generation indicate whether or not lethal mutations occurred on males x chromosomes If lethal mutation occurred on wild type male – it die C – prevents crossing over & recombination So lethality reside in one chromosome itself C – prevents cross over There is 2 markers B – bar eye Wa for apricot
- 15. recessive mutation can only express if there is homozygosity Attached x – to screen F1 mutations on x of males X^X – REPRESENTATION Compound chromosome 2 x chromosome attached Inheritance as single unit X^X female ATTACHED X CHROMOSOME