Jermaine Parker BIOL 311-204 REPORT ON MAPPING EXPERIMENT Fall 2011

Introduction This purpose of this laboratory experiment was to determine the genetic mapping of three mutant genes in Drosophila; (y) yellow body color, (cv) the absence of crossveins on wings, and (f) forked bristles. The recombinant frequencies were used to place the genes relative to each other. After virgin mutant female flies were crossed with wild type males, a F1 generation of wild type females and mutant males was produced. The offspring from the F1 cross were mated with each other, creating the F2 cross. A total of 363 of the F2 offspring were scored to determine the mutant gene locations of y cv and f. Description of the Mutant Phenotypes The first mutant phenotype scored was (y) yellow body color. A mutant fly s body color is much lighter and yellower than wild types, which are dark yellow to brown in color. When comparing a vial that contains mutant flies to a vial that contains wild type flies the difference is obvious. The next gene scored was (f) forked bristles. When examining a fruit fly under a microscope there are hairs on the upper part of the fly s back. On the mutant fly the hairs are stubby and forked at the end, compared to the long unforked bristles of the wild type. The last mutant gene scored was (cv) the absence of crossveins on the fly s wings. Both mutant and wild type flies have long vertical veins that run through the wings; but wild type flies have short horizontal veins connecting some of those vertical ones. Mutant flies do not have horizontal veins crossing over the vertical veins. Segregation Ratios Null Hypothesis: The expected segregation ratio of the two alleles for the body color gene is 1 light brown (y+) : 1 yellow (y). (y) Yellow Body 174 181.5 0.31 (y+) Brown body 189 181.5 0.31 Total 363 363 0.62 = Chi square value

Observed Expected (O-E)2 /E

Degrees of freedom: 1 Probability between: 0.30 and 0.50 Decision: accept the null hypothesis

Null Hypothesis: The expected segregation ratio of the two alleles for the crossvein gene is 1 crossvein (cv+) : 1 no crossveins (cv). (cv) No crossveins 168 181.5 1.00 (cv+) Crossveins 195 181.5 1.00 Total 363 363 2.01= Chi square value

Observed Expected (O-E)2 /E

Degrees of freedom: 1 Probability between: 0.10 and 0.20 Decision: accept the null hypothesis

Null Hypothesis: The expected segregation ratio of the two alleles for the bristle gene is 1 unforked (f+) : 1 forked (f). (f) Forked bristles 171 181.5 0.61 (f+) Unforked bristles Total 192 363 181.5 363 0.61 1.22= Chi square value

Observed Expected (O-E)2 /E

Degrees of freedom: 1 Probability between: 0.20 and 0.30 Decision: accept the null hypothesis In all cases, the null hypotheses were accepted, meaning that the expected segregation ratios of y+:y, cv+:cv, and f+:f are all 1:1. Evaluation of F1 The parental phenotypes were mutant females and wild type males. The genotype of the mutant females was y cv f / y cv f and the genotype of the wild type males was y+ cv+ f+ / Y. The female is homozygous mutant, therefore had a yellow body, no crossveins on its wings, and forked bristles on its back. The wild type males had a darker brown body, the presence of crossveins on its wings, and long unforked bristles on its back. When these two genotypes were crossed, the expected offspring was wild type females and mutant males. This held true in the collection of the FI raw data. My F1 results showed only wild type females (51) and mutant males (21). The number of females collected was larger than the number of males scored for each member of the group. This is probably because mutant male flies take longer to develop. Females in an F1 generation develop faster.

Calculations and Map Construction The gene order is y cv f Phenotype Total Combine (M + F) 110 90 18 16 39 40 27 23 363

+++ y cv f y +f + cv + y cv + ++f y ++ + cv f Total

Non Recombinant Double Crossovers Single crossover Between cv-f Single crossover Between y-cv

Table 4a. Map calculation for y y++ + cv f y+f + cv + Total 27 23 18 16 84

cv pairs (all y+ cv and y cv+ data)

y y

Total y-cv data/ total number of files counted * 100 = map units between y and cv (84/ 363) *100% = 23.14% = 23.14 m.u.

Table 4b. Map Calculation for cv y cv + ++f y+f + cv + Total 39 40 18 16 113

f pairs (all cv+ f and cv f+ data)

y y

Total cv-f data/ total number of files counted *100= map units between cv and f (113/363) *100% = 31.13% = 31.13 m.u f pairs (all y+ f and y f+ data)

Table 4c. Map calculation for y y cv + ++f y ++ + cv f Total 39 40 27 23 129

y y

Total y-f data/ total number of flied counted *100 = map units between y and f. (129/363) *100% = 35.54% = 35.54 m. u. cv and

Since distance between y f is greater than the distance between y between f cv, then y f are on the ends, and cv is in the middle. UNCORRECTED map distance between y and f is 35.54 m.u. y-----------------cv------------------------------f 23.14 m.u. 31.13 m.u.

y---------35.54 m.u.----------f My concern here is that the y cv distance and the cv-f distance do not add up to the total y f distance. In this case, the y f distance does not include the correction for double crossovers.

Table 5. Correction for double crossovers and corrected map. y +f 18 *2 = 36 + cv + 16 *2 = 32 y cv + 39 ++f 40 y ++ 27 + cv f 23 Total 197 (197/363) *100% = 54.27% = 54.27 m.u. CORRECTED map distance between y and f is 54.27 m.u. y----------cv------------------------f 23.14 m.u. 31.13 m.u

y-------------54.27 m.u.----------f I feel much better about this map. The y cv distance and the cv f distance add up to the y f distance, as I expected. This map includes the double crossover corrections. PUBLISHED MAP y---------------cv-------------------------f 13.70 m.u. 43.00 m.u.

y-----------------56.7 m.u.-------------f Discussion The published map unit is displayed above. It shows that the y f distance is 56.7 map units. The corrected map shows that the y f map distance is 54.27 map units. These numbers are very close (only about a 2.4-m.u. difference), and suggest that the experiment was conducted properly. The uncorrected map was off about 19 map units for y-f in comparision to the corrected value. This is because the double crossovers were not accounted for in the uncorrected map. The reasons the data is some points off from the published map units could be due to the sample size and the way the flies were scored. The gender scored for the overall group was not considered but for my individual data, there were more females than males scored. This seemed to be the trend with the F1 data as well, but is not considered as a factor in this genetic mapping experiment. Overall, the experiment showed the proper gene order of the genes being tested.

Experimental Notes My partner and I ran behind schedule at the start of the experiment due to us crossing the wrong types of flies. We were not getting any mutant flies at first. So, around week 3 of the experiment, we restarted the entire experiment from scratch. We quickly caught up. But, my F2 flies were either dying, or I had somehow made a mistake in crossing the F1 generation flies. So, I had to score the remainder of my partner s F2 flies. Everything worked out in the end.

Data tables Table 1. Group F1 Data

Phenotype +++ y f cv Other (if any) Total

My Data Male/Female 51 21

Ahsan s Data Male/Female 24 21

Total Combine (M + F) 75 42

72

45

117

Table 2. Group F2 Data Phenotype +++ y f cv y f+ + + cv y + cv +f+ y ++ + f cv Total My Data Male/Female 75 65 6 8 22 18 20 17 231 Ahsan s Data Male/Female 35 25 12 8 17 22 7 6 132 Total Combine (M + F) 110 90 18 16 39 40 27 23 363