Introduction Genes are unique segments of DNA and in this laboratory experiment, the Actin gene of Drosophila melanogaster will be extracted and amplified with various laboratory processes including PCR, ligation, and transformation. Also, the gene that was extracted will be confirmed and sequenced with the process of cycle sequencing and with the help of NCBI database. The DNA that is first extracted will be referred to as “genomic DNA” because it was extracted directly from the fruit fly, but later on, it will be referred to as “plasmid DNA” and this is when it is incorporated in the plasmid. Drosophila melanogaster or the fruit fly is one of the most commonly used organisms in genetic experiment. They are commonly used because of their small size, four homologous pair of chromosomes, easy maintenance, and easily observable traits (Pierce, 2012).
Groups extracted either the 18S or the Actin gene of Drosophila melanogaster. The 18S gene is located on the flies X chromosome and has a length of 488 base pairs. The Actin gene is located on chromosome 3 of the fly and it has a length of 4,760 base pairs (Adams, 2000), but we are
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In this case, our plasmid DNA is isolated from a liquid culture of the E.coli that was transformed. This is done by reacting the liquid culture with five buffers. The buffers are P1, P2, N3, PE, and an elution buffer. P1 is used to re-suspend the pellet and degrade RNA, P2 is used to lyse cell membrane, PE is used to wash the sample, the elution buffer is used to release DNA from the spin column, and N3 is used to precipitate proteins and genomic DNA. The main components of P1 are Tris, EDTA, and RNase. The main components of P2 are NaOH and SDS. The main component of PE is ethanol and the main component of N3 is acetic acid. The main component elution buffer is water. The possible contaminations of mini-prep are proteins and salts (Garey et al.,
The goal of this study was to induce a deletion in the DMAP1 gene on chromosome two in Drosophila melanogaster through P-element mobilization. The DMAP1 gene may be an essential gene, however not much is known about it. We attempted to uncover the function of DMAP1 by creating a series of genetic crosses and selecting for brown-eyed non-stubble male flies that may have the deletion. To test whether these flies had the deletion, we produced PCR products and ran them on an agarose gel, which resulted as inconclusive. We created a balanced stock of flies homozygous for the deletion to see if the
MacKay JO, Soanes KH, Srivastava A, Simmonds A, Brook WJ, Bell JB. An in vivo analysis of the vestigial gene in Drosophila melanogaster defines the domains required for Vg function. Genetics. 2003;163(4):1365-1373.
When examining the D. Melanogaster mutants in the lab, our group immediately noticed an apparent difference from the wild-type flies. None of the mutants were able to fly. This led us to believe that we were dealing with a wing mutation. Upon further examination, we concluded that it was the overall wing shape that prevented the mutants from flying. The wing shapes among the mutants varied in both size and shape. Some were long, while others were short. The mutant wings could be distinguished into two general classes. One division of the mutant wing was short and stubby, almost a fourth the size of the wild-type wing. The rest of the mutants ranged in wing size and length, however many mutant wings were the same length as the wild-type wings. Although the
Five specific genes are cv, cv-d, cv-c, cv-2, and Dhc16F. cv is named crossveinless and is located at 1-13.7. It is a BMP binding protein that can repress BMP signaling. cv-d is named crossveinless d and located at 3-65. This binding protein decreases the amount of BMP signaling. cv-c is named crossveinless c and is located at 3-54.1. It is a necessary protein for BMP signaling and transport. cv-2 is named crossveinless 2 and is located at 2-96.2. It is a BMP binding protein that increases and decreases the effect of BMP signaling. All of these genes were chosen because they affect the presence of the anterior and posterior crossveins on the Drosophila melanogaster wing. Dhc26F is named Dynein heavy chain at 16F. It is located at 1-59.1. Dhc26F is a motor protein that is used to convert ATP into energy. It was chosen because it consists of the phenotype of a crossveinless-like gene that also affects the presence of the anterior and posterior crossveins on the Drosophila melanogaster
The fruit fly (Drosophila melanogaster) is the model organism when it comes to studying genetics; for over 100 years, it has been used as one of the few model organisms in the scientific community (Twyman, 2002). The reason behind this and why scientists continue to choose to study Drosophila is because of the amount of scientists that have done so and it is well documented on how to treat a fly in a laboratory setting (Twyman, 2002). They are also small and easy to feed. This means that it would be easy to work with a large amount of them without being concerned about taking up too much space, food, or
Sturtevant, A. H. 1913. The linear arrangement of six sex-linked factors in Drosophila, as shown
The F2 Drosophila produced from crossing the wild-type offspring of the parental cross wild-type males and no-winged females. The F2 generation consisted of 38 wild-type females and 35 wild-type males, totaling 73 Drosophila. There were also 16 no-winged females and 11 no-winged males, totaling 27 Drosophila. Therefore, the total number of Drosophila counted was 100. The phenotypic ratio was 3:1, wild-type: mutant. If the mutation were autosomal recessive the F2 generation’s hypothesized phenotypic ratio would be 3:1, wild-type: mutant. If the mutation were autosomal dominant the F2 generation’s hypothesized phenotypic ratio would be 1:3, wild-type: mutant. The expected ratio for the F2 generation was 3:1, wild-type: mutant. When chi-square
Gain of function (ectopic expression in the developing haltere epithelium) of each of these genes changes the haltere of the fly into a second set of wings: double-wings phenotype (GOF).
Scientist use Drosophila melanogaster because they reproduce very rapidly and have shorter generations. These characteristic of being able to go through many generations in a short amount of time combined with its resemblance in behavior and development to a human made it a good candidateto use this organism to study genetics.
S., Boyd, J. B., Carpenter, A. T. C., Green, M. M., Nguyen, T. D., Ripoll, P., Smith, P. D. 1976. Genetic controls of meiotic recombination and somatic metabolism in Drosophila melanogaster. Proceeding National Academy of Science 73(11): 4140-4144.
Plasmids are small DNA atoms that are found in the nucleus region of a bacterium. They provide an extra assistance to a bacterium that allows them to survive and reproduce in certain environments, such as making the bacteria resistant to specific antibiotics. In the experiment being conducted, plasmids pUC18 and lux will be tested to determine the different types of growth with Escherichia coli present. In order to do this, CaCl2 was transferred into a tube with E.coli and incubated for 10 minutes. Different measurements of micro liters of various substances are added to the test tubes and later incubated for 15 minutes. A separate test tube is collected, labeled and added its corresponding micro liters of the competent cells. Once this is
Drosophila melanogaster is a small fruit fly that feeds on fruit and the fungi growing on spoiled fruit. Fruit flies have been used in the research end of the scientific community for over a century due to their interesting physical and behavioral characteristics, their practicality and small size, and their short life cycle of about fourteen days. Its behavior has been the focus of many experiments, beginning with Thomas Hunt Morgan in 1907, and continues today in the laboratories of high school classrooms. In this experiment we are investigating the relationship between a model organism, Drosophila, and its response to different environmental conditions. Our results from the chi-square analysis data all showed signs of our observed fly count
With Drosophila melanogaster being an excellent paradigm for research in gene expression and exploration, many genetic tools have been created in order to utilize the fruit fly’s valuable characteristics in both spatial and temporal control of gene expression. Numerous tools used to study D. Melanogaster derive from the UAS-GAL4 gene expression system, and the system has proven to be quite simple in terms of spatial control of gene expression. When using the UAS-GAL4 system for temporal control, however, things get a little more complicated, and thus a modified version of this system was created. This gene expression system is known as the GeneSwitch GAL4 system, in which an adapted GAL4 protein is fused to a progesterone steroid, and GAL4
After the second Drosophila cross, the phenotypes of 100 F2 Drosophila were scored, rather than 200 phenotypes as described in the GENE222 Laboratory manual. 100 phenotypes were scored because less than 200 Drosophila were produced by the second cross, and 100 phenotypes was a convenient amount for further analysis.
Based on the above mentioned observations, it can be concluded that the quality and genetic diversity of the DGRP (Drosophila Genetic Reference Panel) has made it suitable for an international analyses of transcription factor binding sector variation. It has also been understood that this information is unlikely to elicit a prohibitive bias and that TFBS are functional constraints and can be tolerated so that, possible buffering effects