There are great differences in mating behaviors between humans (Homo sapiens), chimpanzees (Pan troglodytes), and gorillas (Gorilla gorilla). The variability in mating behaviors directly correlates with differences in several aspects of anatomy and physiology among the three species. Much of the variation in semen physiology may be caused by remarkable molecular differences in the length, sequence, and structure of several high abundance proteins of interest. This connection between protein function and physiology could potentially be used to make inferences about the mating behaviors of extinct species, including the common ancestors of humans, chimpanzees, and gorillas. To do this, we will express recombinant proteins from these species in a human cell line (293T cells) and test for functional differences. For the first step of this project, I optimized choice of expression vector, transfection reagents, and transfection conditions using a green fluorescent protein (GFP) expression system. Next, we cloned the KLK3, TGM4, ACPP, SEMG1, and SEMG2 coding sequences of human, chimp, and gorilla into the pFLAG vector, which yielded the best expression during optimization. We detected and quantified expression of these recombinant secreted proteins from media with SDS-PAGE and Western blots. Future work will include purification and functional analysis of these proteins.

Expression of major hominoid seminal proteins in a mammalian cell culture system

249th National Meeting (2015)

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