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Gamble, D.

Publications and source records attributed to Gamble, D..

2 recordsLinked to original sources

Future-proofing code: Principles of coding for plant ecophysiology with {photosynthesis} as a case study

Plant ecophysiology is founded on a rich body of physical and chemical theory, but it is challenging to connect theory with data in unambiguous, analytically rigorous, and reproducible ways. Custom scripts written in computer programming languages (coding) enable plant ecophysiologists to model plant processes and fit models to data reproducibly using advanced statistical techniques. Since many ecophysiologists lack formal programming education, we have yet to adopt a unified set of coding principles and standards that could make coding easier to learn, use, and modify. We identify eight principles to help in plant ecophysiologists without much programming experience to write resilient code: 1) standardized nomenclature, 2) consistency in style, 3) increased modularity/extensibility for easier editing and understanding, 4) code scalability for application to large datasets, 5) documented contingencies for code maintenance, 6) documentation to facilitate user understanding; 7) extensive tutorials, and 8) unit testing. We illustrate these principles using a new R package, {photosynthesis}, which provides a set of analytical and simulation tools for plant ecophysiology. Our goal with these principles is to advance scientific discovery in plant ecophysiology by making it easier to use code for simulation and data analysis, reproduce results, and rapidly incorporate new biological understanding and analytical tools.

plant biology

Recombinational repair of nuclease-generated mitotic double-strand breaks with different end structures in yeast

Mitotic recombination is the predominant mechanism for repairing double-strand breaks in Saccharomyces cerevisiae. Current recombination models are largely based on studies utilizing the enzyme I-SceI or HO to create a site-specific break, each of which generates broken ends with 3 overhangs. In this study sequence-diverged ectopic substrates were used to assess whether the frequent Pol {delta}-mediated removal of a mismatch 8 nucleotides from a 3 end affects recombination outcomes and whether the presence of a 3 versus 5 overhang at the break site alters outcomes. Recombination outcomes monitored were the distributions of recombination products into crossovers versus noncrossovers, and the position/length of transferred sequence (heteroduplex DNA) in noncrossover products. A terminal mismatch that was 22 nucleotides from the 3 end was rarely removed and the greater distance from the end did not affect recombination outcomes. To determine whether the recombinational repair of breaks with 3 versus 5 overhangs differs, we compared the well-studied 3 overhang created by I-SceI to a 5 overhang created by a ZFN (Zinc Finger Nuclease). Initiation with the ZFN yielded more recombinants, consistent with more efficient cleavage and potentially faster repair rate relative to I-SceI. While there were proportionally more COs among ZFN-than I-SceI-initiated events, NCOs in the two systems were indistinguishable in terms of the extent of strand transfer. These data demonstrate that the method of DSB induction and the resulting differences in end polarity have little effect on mitotic recombination outcomes despite potential differences in repair rate.

genetics