LSM 1102 (MOLECULAR GENETICS) Practical 5: DROSOPHILA GENETICS Examination of wild type flies You will be provided with

a small food vial containing male and female wild type Drosophila melanogaster. Etherize (anaesthesize) the flies (with _?_[find out from the lab officers]_) and place them on the white counting plate and note the following characters:Compound eye: Note colour and shape. Ocelli (i.e., simple eyes): These are three single ommatidia on top of the head. Note arrangement and colour. Antennae: Note shape and branches on aristae. Thoraxic and abdominal bristles: Note size, location of bristles and shape. Wing: Note shape, length, position at rest and colour. Note also the pair of balancers or halteres.

How to distinguish the sexes Refer to the figure of male and female D. melanogaster. (below) The female abdomen curves to a point posteriorly and has separate dark bands on the dorsal surface of the abdomen to the very tip. At the tip of the ventral side of the abdomen, the female genitalia (vaginal plate and anal plate) is rather pale in colour. The male is smaller in size and his abdomen has a rounded end with dark bands on the dorsal side of the last few segments fused. On the ventral side of the abdomen, the male genitalia appears as a darkly pigmented area. Examine (with high power) the first tarsal segment of the front legs. Look for a dark hook-shaped structure called the "sex-comb". Is it present in the female?

Life cycle and sexual dimorphism of the vineger fly, Drosophila melanogaster. The male (1) is generally smaller than the female (2). The male also has more rounded abdomen than that is black0tipped rather than striped. The enlarged foreley of the male reveals special bristles known as the sex comb, a characteristic lacking in the female. Metamorphosis is complete, from egg (3), larval stages (4, 5, and 6), pupa (7), through adult.

Examination of some mutants of Drosophila melanogaster WILD TYPE Normal flies with red eyes and other standard characteristics. Mutant flies are inherited departures from this standard phenotype. Wild type means normal and is designated by +. Mutants on the other hand are designated by alphabetical letters. Recessive mutants are designed by small letters (eg. w - white eye) and dominant mutants by capitalising the first letter of the symbol (eg. Cy - Curly wing). Examine the mutants. Make notes of how they differ from the wild type. If in doubt, examine mutant and wild type individuals side by side. Figure 2. Examination of wild type characters in wild-type Drosophila

Figure 3. Some mutant wing types in Drosphila melanogaster

Figure 4. Wing mutants of Drosphila (a) vestigial, (b) curly wings (c) bithorax, (d) dichaete

NAMING OF MUTANTS IN DROSOPHILA The symbol for each mutant is represented by one, two or three letters and its wild-type allele by +. The numbers in parenthesis represent the number of the chromosome carrying the mutation and its linkage location on the chromosome. Small letters are used for recessive mutants and capital letters for the dominant mutants. For example: e = ebony body (3-70.7) e+ = normal body colour Cy = curly wings (2) Cy+ = normal wings svn = shaven bristles svn+ = normal bristles

WING MUTANTS dp : dumpy (2-13.0) Wings reduced to two thirds normal length giving a truncated and cut-off appearance. Thoracic bristles and hair appear unevenly arranged . vg : vestigial (2-67.0) Wings and balancers are greatly reduced. Wr : Wrinkled (3-46.0) Wings do not fully expand after adult emergence but remain crumpled or pebbled and bluish grey in colour and spread apart from each other. The effect is suppressed by Dichaete, completely in the male and almost completely in the female.

BODY COLOUR MUTANTS e : ebony (3-70.7) Body colour gradually turns black in the adult. Larvae show darkened spiracle sheaths compared to the wild-type. y : yellow (1-0.0) The body colour is yellow, with body hairs and bristles brown with yellow tips. The wing hairs and veins are yellow. The larvae mouth parts are yellow to brown, hence are indistinguishable in the larvae stage.

EYE COLOUR MUTANTS + : Wild type. Flies with red eyes and other normal standard characteristics. Mutant types are inherited depatures from this standard phenotype. se : sepia (3-26.0) Brownish red colour which darkens to sepia and finally to black. The ocelli are black.

BRISTLE MUTANTS sn : singed (1-21.0) Bristles and hairs are curled and twisted. Female is sterile.

MULTIPLE MUTANTS cm.ct cm : carmine (1-18.9) Dark ruby eye colour. ct : cut (1-20.0) Scalloped and pointed wings, with reduced eyes and abnormal banding and antennae. w.m.f w : white (1-1.5) White eye colour and colorless ocelli, Malphighian tubules and testes. m : miniature (1-36.1) The wing is reduced in overall dimensions so that it barely extends beyond the tip of the abdomen. The wing surface appears dark because of crowding of cells and hairs. f : forked (1.56.7) Bristles and hairs are shortened and sharply bent or split at the tips, especially those at dorsal surfave of the head and thorax. Cy/Pm D/Sb Cy : Curly (2) Associated with the Curly inversion, in (2L0Cy, on the left arm of the second chromosome. Wings are strongly curled upward and forward. The homozygotes are lethal or dwarfed. Pm : Plum (2- associated woth an inversion). Purplish eye colour, spotted or flecked with darker spots. The homozygote is lethal. D : Dichaete (3-40.4) Wings are held out from the sides at an angle of 45o. The allulae (small basal wing cells) are missing, the balancers are usually turned downward and the dorsocentral bristles are reduced in size. The homozygote is lethal. Sb : Stubble (3-58.2) Bristles are thickened and shortened to less than one half of the usual length. The homozygote is lethal.

Figure 5. Abbreviated linkage map of Drosophila melanogaster (See next page)

Abbreviated linkage map of Drosophila melanogaster. The correlation of each linkage group with one of the four pairs of chromosomes (circular inset) is shown by the dashed lines. Map positions are given on the left of each linkage map; descriptions of the phenotypes produced by the mutant alleles and the genetic symbols used to represent the mutant alleles are given on the right of each linkage map.

DROSOPHILA GENETICS - Take home tutorial please attempt the problems and discuss with your tutors TUTORIAL PROBLEMS Problem 1 Cross a dumpy winged Drosophila with an ebony body fly. Diagram a genetic model to show phenotypes, genotypes of the parental, F1 and F2 generations and their expected ratios. dp: dumpy wing, recessive (2-13.0) dp+: normal wing e : ebony body (3-70.7) e+ : normal body colour

Problem 2 Cross a dumpy winged, ebony bodied fly with a fly with shaven bristles. Diagram a genetic model to show phenotypes, genotypes of the parental and F1 generations and their expected ratios. The F1 was testcrossed to a homozygous recessive tester-stock (dpdp ee svnsvn). Diagram this testcross. dp: dumpy wing, autosomal recessive (2-13.0) dp+: normal wing e : ebony body, autosomal recessive (3-70.0) e+ : normal body colour scn: shaven bristles, autosomal recessive (4-3.0) scn+: normal bristles bristles easpecially those on the abdomen are greatly reduced.

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Problem 3 Cross a male dumpy winged fly with a female white-eyed fly. Diagram a genetic model to show phenotypes, genotypes of the parental, F1 and F2 generations and their expected ratios. dp: dumpy wing, recessive (2-13.0) dp+: normal wing w : white eye, sex-linked recessive (1-1.5) w+ : normal red eye colour

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Problem 4 Cross a female fly with dumpy wings and white eyes to a wild type male with normal wings and red eye colour. Diagram a genetic model to show phenotypes, genotypes of the parental, F1 and F2 generations and their expected ratios. dp : dumpy wing, recessive (2-13.0) dp+: normal wing w : white eye, sex-linked recessive (1-1.5) w+ : normal red eye colour

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Problem 5 Cross a male dumpy winged, ebony bodied fly with a female fly with white eyes. Diagram a genetic model to show phenotypes, genotypes and sex of the parental and F1 generations and their expected ratios. The F1 male was testcrossed to a homozygous recessive testerstock (dpdp ee ww). Diagram this testcross and the resulting progeny. dp: dumpy wing, autosomal recessive (2-13.0) dp+: normal wing e : ebony body, autosomal recessive (3-70.0) e+ : normal body colour w : white eye, sex-linked recessive (1-1.5) w+ : normal red eye colour

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Problem 6 For each of the terms in the left column, choose the best matching phrase in the right column a. phenotype b. alleles c. independent assortment d. gametes e. gene f. segregation g. heterozygote h. dominant i. F1 j. testcross k. genotype l. recessive m. dihybrid cross n. homozygote 1. having 2 identical genes of a given gene 2. the allele expressed in the phenotype of heterozygotes 3. alternate forms of a gene 4. observable characteristics 5. a cross between individuals both homozygous for two genes 6. alleles of one gene separate into gametes randomly with respect to alleles of other genes 7. reproductive cells containing only one copy of each gene 8. the allele that does not contribute to the phenotype of the heterozygote 9. the cross of an individual of ambiguous genotype with a homozygous recessive individual 10. an individual with two different alleles of a gene. 11. the heritable entity that determines a characteristic 12. the alleles an individual has of a gene 13. the separation of the two alleles of a gene into different gametes 14. offspring of the P generation

Problem 7 Describe the charcteristics of the garden pea that made it a good organism of Mendels analysis of the basic principles of inheritance. Evaluate how easy or difficult it would be to make a similar study of inheritance in humans by considering the same attributes you described for the pea.

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Problem 8 In a particular population of mice, certain individuals display a phenotype called short tail, which is inherited as a dominant trait. Some individuals display a recessive trait called dilute, which affects coat colour. Which of these traits would be easier to eliminate from the population by selective breeding (choosing certain individuals to be the parents)? Why?

Problem 9 How many genetically different eggs could be formed by women with the following genotypes? Aa bb CC DD AA Bb Cc dd Aa Bb cc Dd Aa Bb Cc Dd

Problem 10 What is the probability of producing a child that will phenotypically resemble either one of the two parents? Aa Bb Cc Dd aa bb cc dd aa bb cc dd aa bb cc dd Aa Bb Cc Dd Aa Bb Cc Dd aa bb cc dd aa bb cc dd Problem 11 Explain how each of the following phenomena can disrupt Mendelian phenotypic ratios. lethal genes multiple alleles incomplete dominance codominance sex-linkage epistasis

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