Abstract
Drosophila melanogaster is one of the most widely used genetic model systems in biology. The ease of working in an invertebrate model system allows the design and execution of many experiments that would be infeasible in a vertebrate model. Although the strength of the fly as a model system lies primarily in the ease of genetic manipulation, it is flexible enough that biochemical and proteomic approaches can also be used to build a more comprehensive study. Here we present a pair of complementary protocols that we have used to examine sirtuin biology in Drosophila. First, we describe our protocol for measuring lifespan in flies expressing a gene of interest under the inducible control of the Gene-Switch system. Finally, we describe a method for performing chromatin immunoprecipitation on adult flies, including some of the difficulties associated with using this technique in chitinous tissue.
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References
Chippindale AK, Leroi M, Kim SB et al (1993) Phenotypic plasticity and selection in Drosophila life-history evolution. I. Nutrition and the cost of reproduction. J Evol Biol 6:171–193
Helfand SL, Rogina B (2003) From genes to aging in Drosophila. Adv Genet 49:67–109
Tatar M (2011) The plate half-full: status of research on the mechanisms of dietary restriction in Drosophila melanogaster. Exp Gerontol 46:363–368
Duffy JB (2002) GAL4 system in Drosophila: a fly geneticist’s Swiss army knife. Genesis 34:1–15
Xie HB, Golic KG (2004) Gene deletions by ends-in targeting in Drosophila melanogaster. Genetics 168:1477–1489
Gong WJ, Golic KG (2003) Ends-out, or replacement, gene targeting in Drosophila. Proc Natl Acad Sci USA 100:2556–2561
Gong WJ, Golic KG (2004) Genomic deletions of the Drosophila melanogaster Hsp70 genes. Genetics 168:1467–1476
Brand AH, Perrimon N (1993) Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development 118:401–415
Roman G, Endo K, Zong L et al (2001) P[Switch], a system for spatial and temporal control of gene expression in Drosophila melanogaster. Proc Natl Acad Sci USA 98: 12602–12607
Osterwalder T, Yoon KS, White BH et al (2001) A conditional tissue-specific transgene expression system using inducible GAL4. Proc Natl Acad Sci USA 98:12596–12601
Tatar M (2007) Diet restriction in Drosophila melanogaster. Design and analysis. Interdiscip Top Gerontol 35:115–136
Bauer JH, Nur S, Morris S et al (2009) dSir2 and Dmp53 interact to mediate aspects of CR—dependent life span extension in D. melanogaster. Aging 1:1–11
Alekseyenko AA, Larschan E, Lai WR et al (2006) High-resolution ChIP-chip analysis reveals that the Drosophila MSL complex selectively identifies active genes on the male X chromosome. Genes Dev 20:848–857
Larschan E, Alekseyenko AA, Gortchakov AA et al (2007) MSL complex is attracted to genes marked by H3K36 trimethylation using a sequence-independent mechanism. Mol Cell 28:121–133
Negre N, Lavrov S, Hennetin J et al (2006) Mapping the distribution of chromatin proteins by ChIP on chip. Methods Enzymol 410:316–341
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Wood, J.G., Whitaker, R., Helfand, S.L. (2013). Genetic and Biochemical Tools for Investigating Sirtuin Function in Drosophila melanogaster . In: Hirschey, M. (eds) Sirtuins. Methods in Molecular Biology, vol 1077. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-637-5_4
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DOI: https://doi.org/10.1007/978-1-62703-637-5_4
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-636-8
Online ISBN: 978-1-62703-637-5
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