Introduction The relatively short generation time of drosophila melanogaster in addition to the geometric increase in their population number

makes the species an ideal organism for studying inheritance patterns of genetic traits. The following experiment took three of those traits under considerations in an effort to ascertain the dominance/recessive, sex-linked/autosomal status of those traits. The three traits that were examined in the following experiment were the wing size, bristle shape, and eye color. Long wings, long bristles, and red eyes were the wild type traits. Conversely, short wings, arched bristles, and blanched eyes were the mutant traits. These were the fundamental assumptions that the experimenters used in order to guide the course of the entire experiment. Wild type and mutant traits fall under the larger paradigm of dominant and recessive traits. Dominant traits are those traits that are expressed in the presence of only one allele of the gene. The one allele is sufficient to produce the biochemical product responsible for the expression of the gene. Recessive traits can be better thought of as the lack of expression of the trait. The dominant allele is not present on either strand of the chromosome, and thus, the biochemical product responsible for the expression of the gene is not produced. By their inherent virtue, individuals possessing recessive traits are homozygous, that is, they are pure lines. Essentially, this means that they possess identical alleles of a gene on the two strands of their chromosome. However, the genotypes of individuals possessing dominant traits cannot be as readily discerned. Again, this is due to the fact that only one dominant allele is required to produce the dominant phenotype. The genotype of the individual possessing dominant characteristics can only be identified with certainty by utilizing a test cross, in which the dominant individual is

crossed with a recessive individual and the phenotypes of the progeny is examined. Certain ratios of progeny characteristics will shed light on the genotype of the dominant parent. If all the progeny display the dominant characteristic, then the dominant parent is homozygous dominant, or a pure line. If half of the progeny display the dominant characteristic and half of the progeny display the recessive characteristic, then the dominant parent is heterozygous dominant, meaning that she/he possesses both the dominant allele and the recessive allele for a given trait. In addition to understanding the dominant/recessive nature of alleles, their behavior can be further understood by examining their location. Genes can either be located on sex chromosomes the chromosomes primarily responsible for gender or they can be located on autosomes all other chromosomes in an organisms genome. Autosomal genes follow the basic Mendelian style of inheritance irrespective of the gender of the parent or progeny. However, gender is a crucial factor in sex-linked inheritance. To our current understanding, all known sexlinked genes lie in the X-chromosome. Thus, a male who possesses a sex-linked dominant trait who mates with a female who is recessive for that trait will produce daughters that will all possess and display that trait because the male has only his one X-chromosome with dominant alleles to pass on to his daughters. However, the sons of such a mating will all be recessive for that trait because the father cannot pass on his X-chromosome to his sons (Griffiths, Miller, Suzuki, Lewontin, & Gelbart, 2002, pp.38-41). Once these basic elements of inheritance are understood, one must examine inheritance at a higher level. This entails a thorough analysis of the linkage of certain genes on a chromosome. Linkage is a phenomenon that defies the rules of classical Mendelian inheritance. Essentially, this means that the closer two genes lie to each other on a chromosome, the more likely that the two phenotypes produced by those genes are going to be observed in concert. For example, if

the genes for wings and bristles in Drosophila lie close to each other on a chromosome, then following a test cross of an F1 heterozygous individual, one will observe a predominantly large number of progeny that display the wing and bristle characteristics of the two parental strains. However, characteristics other than those in the original parental strain will also be observed. This is the result of a phenomena referred to as crossover (Griffiths, Miller, Suzuki, Lewontin, & Gelbart, 2002, pp.142-143). Crossover occurs during meiosis. The initial stages of meiosis differ from those of mitosis in that there are double the numbers of chromosomes in prophase I of meiosis than in prophase of mitosis. These homologous chromosomes both contain two sister chromatids. Each homolog is inherited from each parent. During prophase I, chromatids of homologous chromosomes are criss-crossed at numerous places. These locations are referred to as chiasmata (Campbell, Reece, & Mitchell, 1999, pp. 232-233). These chiasmata are the visible manifestations of crossovers (Griffiths, Miller, Suzuki, Lewontin, & Gelbart, 2002, p. 143). It is during crossovers when homologous chromosomes exchange parts, and yield a new set of chromosomes that are different in allelic composition than either of the parental chromosomes. The application of the cross-over process allows us to map genes on chromosomes. Mapping is based on Sturtevants postulate: the greater the distance between linked genes, the greater the chance that non-sister chromatids of homologous chromosomes will cross over in the region between the genes(Griffiths, Miller, Suzuki, Lewontin, & Gelbart, 2002, p.147). The recombination frequency (RF) obtained from a mapping studying directly translates into the distance between two genes, or genetic map unit (m.u.). When forming genetic maps, we must also take into consideration the number of double recombinants, or double cross-overs that occur during meiosis. The two rarest classes in a three-

point testcross represent the instances of double recombination. The total of these two classes represent the observed number of double recombinations. The expected number of recombinations can only be calculated once the gene order has been identified. Then, the product of the two adjacent RF values is multiplied by the total number of individuals in order to obtain the expected number of recombinations. The proportion of observed frequency of recombination to expected frequency of recombination is referred to as the coefficient of coincidence (c.o.c.). This number subtracted from 1 reveals the interference value. Thus, the interference value basically measures the likelihood of single cross-overs (Griffiths, Miller, Suzuki, Lewontin, & Gelbart, 2002, pp.151-152). Methods September 9, 2003: the instructor, Professor Frey, performed all the etherization procedures of the experiment. In order to etherize, one must be careful not to over/under-etherize the container. Etherization is carried out by placing two squirts of ether inside the bottom aspect of an etherizing container. The pre-etherized mutant and wild-type flies were obtained from their respective containers. The sex of the flies was determined under a dissecting scope. The objective was to obtain 2 mutant females and 1 wild type male. The following 3 flies were identified and designated as the parental (P) generation. 2 females: short wings (+); blanched eyes (+); arched bristles (+); 1 male: long wings (L); red eyes (A); long bristles (B). The 3 flies were then placed inside a container with drosophila media provided by Carolina Biological Supply, formula 4-24 where they were allowed to mate.

September 23, 2003: The container, which now contained the F1 Generation, was obtained from Professor Frey. The container was opened outside the Science & Technology Building of Indiana University Purdue University Indianapolis, where the adult flies were allowed to escape. The larvae were retained. September 25, 2003: The container, which now contained F1 virgins, was obtained again from Professor Frey. 12 mutant males were retained. 12 wild type females were obtained from Professor Frey. 2 containers of Carolina Biological Supply Drosophila media formula 4-24 were filled with 6 mutant males and 6 wild type females each. October 14, 2003: The containers, which now contained the F2 Generation, were obtained from Professor Frey. Two squirts of ether were placed inside two etherizing containers each. Each container with the F2 generation was placed over each container with the ether. This procedure was done with a certain degree of alacrity in order to prevent loss of F2 flies. Once the flies were sedated, they were examined under a dissecting scope. The gender of the flies and the number of flies was tabulated according to these characteristics. LAB| +++ | L++ | +AB | L+B | +A+ | LA+ | ++B. Results The results of our groups tabulation of Drosophila numbers based on the selected traits are as follows: Phenotype +++ LAB L+B +A+ LA+ ++B L++ +AB Total Male 38 27 10 13 15 19 7 9 138 Female 48 28 13 18 17 22 9 7 162 Total 86 55 23 31 32 41 16 16 300

Table 1: Group-Wise Numbers of Drosophila Phenotypes of F2 Generation The results of the entire laboratorys tabulation of Drosophila numbers based on the selected traits are as follows: Phenotype Male Female Total +++ 745 947 1692 LAB 552 573 1125 L+B 197 240 437 +A+ 240 355 595 LA+ 300 348 648 ++B 351 426 777 L++ 135 150 285 +AB 145 139 284 Total 2665 3178 5843 Table 2: Laboratory-Wise Number of Drosophila Phenotypes of F2 Generation Discussion Our first analysis was derived from the F1 generation. Our P generation consisted of one wild type male (+++) and two mutant females (LAB). All F1 male progeny possessed the phenotypes of their mothers. All F1 female progeny possessed the phenotypes of their father. As it was discussed in the introduction, this is a hallmark characteristic of a sex-linked recessive trait. The data in tables 1 and 2 were used to map the genes on their respective chromosome. Mapping was done on both the group-wise (Table 1) and laboratory-wise (Table 2) data. For the group-wise data, the calculations are as follows: Rf (L and A loci): 23+31+16+16/300 X 100 = 28.7% Rf (L and B loci): 32+41+16+16/300 X 100 = 35.0% Rf (A and B loci): 23+31+32+41/300 X 100 = 42.3% Thus, the resultant sequence of genes on the chromosome is such: A--------------L-------------------B

28.7 m.u.

35.0 m.u.

The observed number of double recombinant classes is 16+16=32. The expected frequency of double recombinants is 0.35 X 0.287 = 0.10045 The expected number of double recombinants is 0.10045 X 300 = 30 The c.o.c. is 32/30 = 1.07 The Interference (I) is 1 1.07 = 0.07. The negative value of I suggests that there is a zero probability of purely single cross-overs of the following genes on the chromosome. By an extension of that outcome, there is a definite chance of double cross-overs of the genes on homologous chromosomes. The large map units between the genes confirm this deduction. For the laboratory-wise data, the calculations are as follows: Rf (L and A loci): 437+595+285+284/5843 X 100 = 27.4% Rf (L and B loci): 648+777+285+284/5843 X 100 = 34.1% Rf (A and B loci): 437+595+648+777/5843 X 100 = 42.1% Thus, the resultant sequence of genes on the chromosome is such: A--------------L------------------B 27.4 m.u. 34.1 m.u.

The observed number of double recombinant classes is 285+284=569 The expected frequency of double recombinants is 0.274 X 0.341 = 0.093434 The expected number of double recombinants is 0.093434 X 5843 = 546 The c.o.c. is 569/546 = 1.04 The Interference (I) is 1 1.04 = 0.04

Similar to the group-wise data, the negative I indicates a definite chance of double cross-overs of the genes on homologous chromosomes. References Griffiths, A.J., Miller, J.H., Suzuki, D.T., Lewontin, R.C., and Gelbart, W.M. (2002). An introduction to genetic analysis (7th ed.). New York: W.H. Freeman and Company. Campbell, N.A., Reece, J.B., & Mitchell, L.G. (1999). Biology (5th ed.). New York: Benjamin Cummings.