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Cutting genes with undergraduate scissors
Author(s) -
Marintcheva Boriana,
Young Kelly,
Smith Crystal,
MacDougall Lawrence,
DaConceicao Silviane,
Tandon Ankita
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.838.9
Subject(s) - plasmid , gene , biology , genome , clone (java method) , genetics , mutant , computational biology , bacteriophage , synthetic biology , escherichia coli
Bacteriophage T7 is a genetically and biochemically amenable model system offering great opportunities to train undergraduates in molecular biology and to empower them to pursue independent projects. Students enrolled in BIO497/Undergraduate Biological Research at Bridgewater State University were able to experience science as a process while engineering deletion mutants of T7 DNA metabolism genes. PCR was used to amplify genes of interest from the T7 genome and to clone them using TOPO technology. The resulting plasmids were transformed in E. coli generating strains permissive for growth of phage deletion mutants. In parallel, multi‐step PCR was used to generate recombination cassettes carrying the sequence for cytidine monokinase ( cmk ), flanked by the natural neighboring sequences of each gene of interest. Cmk is essential for T7 growth, but dispensable for the host, therefore an ideal selection marker for recombinant viruses. In vivo recombination reaction was carried out resulting in the replacement of each individual gene of interest in the viral genome with the cmk sequence. Throughout the experience students learnt multiple techniques, sharpened their problem solving and time management skills, wrote grant proposals, and presented their work locally and nationally. This project was generously supported by the Adrian Tinsley Undergraduate Research Program at Bridgewater State University.