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Drosophila melanogaster Materials and Methods

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Drosophila melanogaster Materials and Methods

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Materials and Methods

The common fruit fly or vinegar fly belongs to the family of Drosophilidae is one of the most used model organisms for biomedical science. According to Markow and O’Grady (2005) its current distribution is worldwide, being found on every continent and most islands.  Two strains of fruit flies of the species Drosophila melanogaster were obtained. One strain consisted of true-breeding wild-type flies and the other consisted of true-breeding flies with two mutations, the dominant Wrinkled (Wr) mutation and the recessive black (b) mutation. Standard methods were used in anesthetizing and sexing the flies (Galbraith, et al., 2008). Mutant virgin female flies were acquired as described (Flagg, 2001) and two of these mutant virgin females were placed into each of two vials along with two wild-type male flies. Vials were set up containing a potato flake based medium as described in Flagg (2001).

After about 7 days, the vials were checked for F1 larvae. As soon as F1 larvae were apparent, the parent flies were anesthetized and discarded to prevent backcrossing.  Once the F1 larvae had pupated and hatched into adult flies, 3 vials were set up, each containing two F1 females and two F1 males. The flies involved in theF1 cross were allowed to mate for about one week until F2 larvae were observed in the vials. Once F2 larvae were present, the F1 parents were anesthetized and removed from the vials.

After the F2 larvae pupated, the vials were checked daily to note the first day of hatching in each vial. F2 flies were counted and scored every three days for 12 days. After scoring, the flies were discarded. A chi square analysis was performed on the data to determine whether the observed numbers were statistically similar to expected results.

A BLASTn search (NCBI, 2008) conducted using the wild-type sequences for both the Wrinkled and black genes to identify genes from other organisms with similar sequences. The E values for the alignments between the sequences, and putative or known functions of these genes were used to determine whether the resulting alignments reflected a similarity between the genes that was biologically relevant.   

Most populations of domestic and wild animal and plant species at risk of extinction are spatially structured. This structuring must be explicitly considered in the conservation management of genetic resources and the fields of animal breeding and evolutionary biology. For instance, most species undergoing conservation programs in captivity are generally maintained in independent nuclei  dependent nuclei (zoos, botanic gardens, germplasm centers and so on) (Valladares-Padua et al., 2002Watanabe et al., 2009). In some situations, subdivision has a clear biological meaning, as different subpopulations characterized by local adaptations (Storfer, 1999Branch et al., 2003).

Over a century, due to its many technical advantages including its short life span, of using Drosophila over vertebrate models. They are easy and inexpensive to culture in laboratory conditions, have a much shorter life cycle, produce large numbers of externally laid embryos and can be genetically modified in numerous ways.

Drosophilia has been used as a model organism to study a wide range of biological processes including inheritance and genetics, learning behaviour, embryo development an aging. Similarities between humans and fruit flies biological mechanisms and pathways that control development and survival are uncanny across evolution between these species.

Two strains of fruit flies of the species Drosophila melanogaster were obtained. One strain consisted of true-breeding wild-type flies and the other consisted of true-breeding flies with two mutations, the dominant Wrinkled (Wr) mutation and the recessive black (b) mutation. Standard methods were used in anesthetizing and sexing the flies (Galbraith, et al., 2008) Mutant virgin female flies were acquired and placed into a vial each  along with two wild-type male flies (Flagg,2001). Flagg (2001) states the vials were set up containing a potato flake based medium .

References

(Science. https://science.sciencemag.org/content/sci/suppl/2003/05/01/300.5620.798.DC1/Naber.SOM.pdf

Markow, T. A., & O’Grady, P. M. (2005). Evolutionary genetics of reproductive behaviorin Drosophila: connecting the dots. Research Gate 39(1). doi:10.1146/annurev.genet.39.0703003.112454

(Galbraith, et al., 2008).

Flagg (2001).

(Storfer, 1999Branch et al., 2003).

) (Valladares-Padua et al., 2002Watanabe et al., 2009)

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