Scientific Research + Laboratory Experiments

A Two-Hybrid Screen to Test C. elegans Heterochronic Gene Lin-66 for Protein-Protein Interactions

This is a research paper I wrote during my junior year using data from original experiments I performed with a lab team in a foundational biology lab. In this experiment, DNA containing a gene of interest from the worm C. elegans was placed into yeast cells and later E. coli. The cells underwent various procedures to test for specific protein-protein interactions.  This paper demonstrates my ability to display, explain, and interpret raw data.  It also illustrates my ability to write a research paper in the style of those in a peer-reviewed journal, such as Nature. Additionally, it displays my understanding of many common scientific procedure and protocols, such as using plasmid vectors to perform cross-species DNA manipulation. Many undergraduate students in my major don’t have this kind of first-hand experience with the scientific experiments they might read or write about. The following excerpt is from the “Abstract” portion of the paper.

Abstract

A yeast two-hybrid screen was used to test a C. elegans cDNA library for protein-protein interaction with the protein LIN-66, encoded by the heterochronic gene, Lin-66. LIN-66 has been shown to play an important role in developmental timing and the developmental mechanisms of C. elegans are useful for study of all animal development, because heterochronic genes are highly conserved across species. From the two-hybrid screen, four potential interacting strains were isolated through selective-nutrition growth tests and an X-Gal filter assay.  A HindIII restriction digest and gel electrophoresis were performed to confirm the identity of these strains as non-Lin-66 DNA fragments.   Of these four strains, only one was confirmed as a non-Lin-66 DNA fragment.  This strain was submitted for DNA sequencing and results were analyzed with a BLAST search. The gene identified is rfl-1, which encodes for a subunit of an enzyme which plays a critical role in the NEDD8 ubiquitin-like conjugation pathway.

Click to download a PDF copy of the entire paper.

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