Bioinformatics uses complex algorithms and computer software to make predictions about genetic material. However since this is done<i> in silico, </i>these predictions still need biological confirmation—a difficult and time-consuming process. The goal of this project is two fold: first, to examine characteristics of the regulatory elements found in mycobacteriophage using novel cloning techniques; second, to investigate the validity of commonly used algorithms to ensure that the data produced from genomic analysis can be properly interpreted. To accomplish this goal, fourteen putative promoters from mycobacteriophage Mendokysei were identified through bioinformatic analysis using DNA Master. Additionally, six terminators were identified using ARNOLD. Promoters were verified and quantified using the expression vector pClone Red (BBa_J119137). A novel construct for terminator analysis named PGR-Blue was developed to increase the efficiency and accuracy of terminator quantification. Both vectors utilize Golden Gate Assembly to allow for the rapid testing and selection of putative regulatory elements. Results show that mycobacteriophage Mendokysei has one strong promoter and several weak promoters; furthermore, results show Mendokysei has two functioning terminator sequences.

Biological characterization of mycobacteriophage promoter and terminator identified through bioinformatics

251st National Meeting (2016)

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