Designing Functional Ripening-Control Tomato Promoters
Tomato, Solanum lycopersicum has been the subject of many studies focused on the development and ripening process of climacteric fruits; studies focus on pigmentation, texture, aroma, flavor, and nutrient composition (Moore et al., 2002). To study fruit ripening, it is necessary to use ripening mutants. Two naturally occurring recessive mutants ripening inhibitor (RIN) belongs to the MADS-box gene family and another mutant non-ripening (NOR) belongs to the NAC family. Both gene are transcription factors that control ripening.
The goal of this internship was to design functional RIN and NOR promoter constructs. Four promoter constructs were designed for NOR and three were designed for RIN to elucidate the most efficient constructs. Promoter constructs are assembled within the pBi101 transformation vector using GUS gene expression assay. Constructs were created with at least 3.2 Kb of promoter sequence upstream of the 5′ translational start site. Promoter regions were paired with ‘required’ intron regions of DNA to include possible cis-regulatory elements within constructs to achieve high expression level. These functional promoters will be used as a tool for genes that need to be expressed in a fruit-specific manner.
Overall, the internship was a great success. I was lucky enough to be in a lab with two other interns. The lab environment was surprisingly laid-back and very easy to integrate into. All the post-docs, research associates, and graduates students were excellent resources for my project and for my growing curiosity in graduate degree programs. Working directly with a mentor on my research was beneficial in learning new scientific techniques and for gaining an understanding of the dedication a scientific career requires. Through my experience and interaction with my peers I was able to gain an accurate view of the career paths available in plant research and glimpse the projects graduate students focus on during the PhD research.