Identification of eukaryotic promoter regulatory elements using nonhomologous random recombination

By یکىجرا یدیما, دیما, Omid Omidi-Arjenaki, -. D. Ghanbarian, -. M. Naderi-Boldaji, -. K. Mollazadeh. Nucleic Acids Research, 2007

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Understanding the regulatory logic of a eukaryotic promoter requires the elucidation of the regulatory elements within that promoter. Current experimental or computational methods to discover regulatory motifs within a promoter can be labor intensive and may miss redundant, unprecedented or weakly activating elements. We have developed an unbiased combinatorial approach to rapidly identify new upstream activating sequences (UASs) in a promoter. This approach couples nonhomologous random recombination with an in vivo screen to efficiently identify UASs and does not rely on preconceived hypotheses about promoter regulation or on similarity to known activating sequences. We validated this method using the unfolded protein response (UPR) in yeast and were able to identify both known and potentially novel UASs involved in the UPR. One of the new UASs discovered using this approach implicates Crz1 as a possible activator of Hac1, a transcription factor involved in the UPR. This method has several advantages over existing methods for UAS discovery including its speed, potential generality, sensitivity and lack of false positives and negatives.

Subfields: Biotechnology, Cell Biology, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/25e188d2465d8fb5e2250288c020432cdb99e789

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In Molecular Biology

Citations in the corpus

Antibiotic resistance plasmids in wastewater treatment plants and their possible dissemination into the environment (2010, methodology): “recently, non-homologous random recombination (NRR) method has been described, allowing recombining nonhomologous proteins, which is difficult or impossible to accomplish through the previous techniques (Bittker et al., 2004; Doyon and Liu, 2007).”