This page of my website is a summary of the projects I'm doing in the biochemical pharmacology department of the University of Buffalo. Give me time and I'll make it more detailed but for now what you see is what you get.
The goal of this project is to create a double mutant Drosophila Melangaster (fruit fly). The mutations are located on the 1st and 3rd chromosomes, there being 4 chromosomes all together. The mutations I am trying to obtain are "ebony" and "apricot eye" on the same fly. These mutations are located in the phenotype of the fly and are characterized by a black exoskeleton (as opposed to the wild type, being a light brownish color), and yellow-orange eye color (as opposed to the wild type, being red eye color).
The flies I have to start with, or rather my first cross, are a DB2 female and an eag^sc29 male. DB2 is a sort of nickname for a female with the mutations of "kidney eye," "ebony," "tubby," and "serate." "Kidney eye" is the mutation that occurs in heterozygous chromosomes containing the FM7 gene. The characteristic in the phenotype only occurs in the female fruit fly and causes the eye to be in the shape of a kidney, which really goes without saying. "Tubby" and "Serate" are both located on the 3rd chromosome and cause the fly to be short and stout, and the wings to have a small piece missing from each. Eag^sc29 is the gene for "apricot eye" and the male fly I am using has only that mutation. From this first cross, my goal is to produce a female fly with only the mutations of "kidney eye" and "serate." This fly I will cross with a 9g male. 9g is the gene that produces "ebony" in flies. There are other genes, such as TM3 (the gene for "serate") and TM6 (the gene for "tubby"), that also cause "ebony" although they are not as prominent. These two new flies will be the start of my second cross.
From this second cross, I wish to obtain another "kidney, serate" female, and a male with the characteristics of "round eye," the wild type which appears as round, red eyes in the phenotype, and "serate." These characteristics are markers for the genes that I must obtain from the fly, which has the eag^sc29 gene although not identifiable by the "apricot eye" that it is associated with due the fact that the gene is recessive. I will need this male fly to start my third and fourth crosses.
The "kidney, serate" female and the "round, serate" male will be used for the third cross, from which I need to obtain a female fly to cross with the second "round, serate" male for the fourth and final cross.
From this last cross I will obtain an "apricot, ebony" male and a female of the same phenotype. I have currently started my third cross.
The process of breeding the flies is slightly more complex than it may seem. The beginning process consists of checking each fly to be certain it has the genes needed and no others. When the flies are crossed, they are put in a vile together for roughly 2 to 4 days in order for the flies to mate and the male to fertilize the eggs in the female. She lays them in the food culture, usually made of yeast. When larva are discovered, the flies are transfered into another vile in order to keep the cross going. The larva left behind develop into pupa, and then hatch into flies. The flies usually hatch around 8am and they must be scored as soon as possible. This is in order to keep the females virgins because any crosses that have non-virgin females will offsprng that is not wanted and will ruin the experiment.
Scoring involves looking at each individual fly under a microscope and finding a fly with the desired characteristics in the phenotype.
The whole process is then repeated the desired amount of times, in my case 4, until the desired flies are obtained, thus completing the experiment.
