Exogenous expression of Scgblcl in understanding its role in cardiac function

Kevin Guerrero

Document Type

Article

Publication Date

Summer 2018

JAX Location

In: Student Reports, Summer 2018, The Jackson Laboratory

Abstract

Secretoglobin proteins have functions in inflammation modulation, tissue repair and tumorigenesis (1). A previous study demonstrated downregulation of Secretoglobin family le member l, Scgbl cl, in Nkx2-5 knockout mice models, which led to possible roles in neonatal and adult cardiac development (2). By overexpressing the gene and supplying purified Scgblcl protein in neonatal cardiomyocytes, we aim to obtain novel information on the cardiomyocyte roles for Scgbl cl in homeostasis and under cellular stress that could lead to a better understanding of this novel protein's mechanism of action. The purpose of this study is to formulate optimal parameters for Scgbl cl expression and purification while continuing the process of identifying a function for Scgbl cl in neonatal to adult cardiac development. Utilizing Rosetta2(DE3) as a host organism resulted in weak expression of Scgbl cl at various temperature. Bl21star(DE3) resulted in good expression of Scgblcl. To improve upon the expression, replacing the mammalian biased codons with E. coli biased codons would likely increase levels of bacterial expression. Additionally, the protocol for transfection in cardiomyocytes may need improvement to increase efficiency. Nonetheless, continuing studies to analyze the gene and understand its physiology will be the focus for future work.

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