Molecular Biology

As a Genetics Instructor at Grand View University, during Spring 2017 I taught an upper-level Molecular Biology laboratory course for biotechnology majors. This course included a semester-long project on isolating and sequencing of the GAPDH gene (the gene for glyceraldehyde-3-phosphate dehydrogenase) from Coriandrum sativum (cilantro) and Salvia rosmarinus (rosemary). These plants were chosen by the students as plant species with no previously published sequence for the GAPDH gene. So it was a great opportunity for the students to not only be trained in standard molecular biology techniques (e.g., DNA extraction, PCR, DNA ligation, transformation, restriction digestion, etc.) but also learn sequence analysis and have an opportunity to deposit a novel sequence in the NCBI GenBank database.

Topics covered

1. Extraction of genomic DNA
2. PCR amplification of a region of the GAPDH gene
3. Purification of the PCR product
4. Ligation of the PCR product into a plasmid vector
5. Bacterial transformation with the plasmid
6. Restriction digestion analysis
7. DNA sequencing
8. Bioinformatics analysis of the cloned gene

Cloning and sequencing of the GAPDH gene from Coriandrum sativum and Salvia rosmarinus: research project with my students

During the semester my students successfully isolated, cloned, and obtained novel partial sequences of the GAPDH gene from both Coriandrum sativum and Salvia rosmarinus. The students presented their results at annual Grand View Scholarship Symposium (April 21, 2017). Also, our GenBank submission for the partial sequence of the GAPDH gene for Salvia rosmarinus were recently approved (GenBank Accession no. MF074139).