Abstract
Cis-regulatory sequences control when, where, and how much genes are transcribed. A better understanding on these elements is a fundamental keystone to better understand development, cell differentiation, and morphogenesis. Several methods based on in silico analysis or ChIP-seq experiments have been developed to detect cis-acting sequences. Here, we describe a protocol to isolate such sequences from genomic DNA and to clone them into expression vectors for functional assays using the Gateway cloning technology.
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Acknowledgments
We would like to thank R. Freitas, E. de la Calle-Mustienes, and J. Tena for comments on the manuscript. We also thank the Gomez-Skarmeta`s laboratory for the development and improvement of protocols. J.B. is a postdoctoral fellow of the Portuguese Fundação para a Ciência e a Tecnologia (POPH/FSE). This work was supported by grants from the Spanish Ministry of Education and Science (BFU2010-14839 and CSD2007-00008), Junta de Andalucía (CVI-3488) to JLG-S and by EFSD/Lilly to JB. CABD is institutionally supported by CSIC, Universidad Pablo de Olavide, and Junta de Andalucia.
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Bessa, J., Gómez-Skarmeta, J.L. (2012). Making Reporter Gene Constructs to Analyze Cis-regulatory Elements. In: Orgogozo, V., Rockman, M. (eds) Molecular Methods for Evolutionary Genetics. Methods in Molecular Biology, vol 772. Humana Press. https://doi.org/10.1007/978-1-61779-228-1_23
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DOI: https://doi.org/10.1007/978-1-61779-228-1_23
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