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The Effects of Thatch Ants on the Pollination of Oval-leaved Milkweeds

An Independent Research Summer Program study conducted by Karen Beck

In this study, the effects of thatch ants on milkweed pollination was studied by excluding ants from the milkweed flowers, observing the ants and their interactions with other pollinators, measuring the amount of nectar in the flowers with and without ants, and measuring the amount of time that flying pollinators spend on the flowers with and without ants.  It was hypothesized that the pollination of A. ovalifolia would be greatest when the thatch ants are excluded from the flowers.  Research was conducted at the University of Minnesota’s Cedar Creek Ecosystem Science Reserve in the summer of 2012.

Cloning the GAPDH encoding gene GAPC from Thymus vulgaris

Class project in Genetics (BIOL 2202) completed by Amanda Wendel, Ann Kopiecki, Angela Eromobor, Uchenna Umerah and under the mentorship of Paula Croonquist

GAPDH gene is considered a housekeeping gene because it codes for an important step of glycolysis, and is found in the genome of all plants. The gDNA of thyme was extracted, and a large amount of GAPDH gene was synthesized using PCR. Cloning procedures included Blunt end ligation with p Jet1.2 clonining vector, transformation of the plasmid into E.coli followed by incubation in a selective medium. In preparation for DNA sequencing, a miniprep of the plasmid DNA was prepared. Verification of insert of interest and determination of sizes of the band using gel electrophoresis of BgL II restriction digest fragments confirmed the presence of the insert of interest.

Discovering the Role of Hyaluronan Synthase 2 in Melanoma

Mercy N. Konchellah (IRSP student); Matthew Price; and James B. McCarthy, Ph.D.
University of Minnesota, Minneapolis, MN

Condensed Abstract: Melanoma is a type of skin cancer that affects the pigment producing cells known as melanocytes, found in the epidermis. Hyaluronan Synthase 2 (HAS 2) belongs to a family of enzymes, hyaluronan Synthases (1-3), responsible for Hyaluronan (HA) biosynthesis. Previous evidence indicates that this protein family contributes to the progression of cancer but in ways that are still under investigation. The aim of this study was to investigate the levels of HAS 2 protein expression and its impact in melanoma. HAS 2 protein expression in several Melanoma cell lines was assayed by Western blotting. In addition, these cell lines were treated with HAS 2 short interference RNA (siRNA) to knock down expression of HAS 2. Migration assays were set up following the inhibition of HAS 2 mRNA in these cells. We found that Melanoma cells overexpressed HAS 2 protein. Furthermore, HAS 2 knockdown cell line A375, a metastatic melanoma cell line, migrated significantly less than control cells. We concluded that HAS 2 deregulation may affect the migration of metastatic melanoma.

GAPC Gene Cloning in Purple Prairie Clover

Katarina Conradson, Shane Turgeon, Nicole Benson, Ian Walbridge, Paula Croonquist

The objective of this experiment was to clone and sequence the GAPDH encoding gene GAPC for sequencing in purple prairie clover. The central question was to investigate whether the GAPC gene is conserved in the Dalea purpurea, as in other plants.  It was hypothesized that the GAPC gene is found in Dalea purpurea, and that although it constitutes a unique sequence not previously reported, it shares a high degree of homology with other cytosolic GAPDH encoding genes in other plant species.  In this experiment, plant cells were lysed, and DNA was extracted, purified, and run through PCR to amplify the GAPC gene.  E. coli was used to clone the gene of interest, which was then extracted.  Gel electrophoresis was utilized twice, after PCR reactions and digesting extracted vectors, to separate and visualize amplified gene product.  The PCR reactions did not yield any PCR products, and previously amplified PCR products were ligated into the vector.  The cloned vectors were extracted from the transformed bacteria and digested to separate the gene insert from the vector.  The electrophoresis gel of the vector digest did contain bands consistent with a successful gene insert.  It was concluded that samples contained the GAPC gene insert, thus indicating successful isolation and cloning of this gene in purple prairie clover.

Stream Suitability and Potential Economic Impact of Trout Introductions to Trott Brook

Independent Study Research conducted by Aurore Phenow.

The North Metro Area of the Twin Cities has no significant trout streams that could attract anglers, and Trott Brook was identified as a potential stream for trout introduction.   Water quality and aquatic macroinvertebrates were sampled in August-October 2012 and physical characteristics were measured to determine the feasibility of converting this section of Trott Brook to a viable trout stream, and a literature review of the economic impact of trout-angling was also conducted.  Many aspects of this stream were positive indicators for trout but monitoring temperature during the summer months would be key to concluding whether Trott Brook could support a sustainable population of trout.

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