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Cloning Isoform 1 of TEM8

Undergraduate #41
Discipline: Biological Sciences
Subcategory: Cell and Molecular Biology

Latisha Franklin - Dillard University
Co-Author(s): Jenn White- Bromberg and Nick Duesbery, Van Andel Research Institute



Peripheral Arterial Disease (PAD) is a circulatory issue caused by narrowed arteries; patients suffer from a reduced ability to expand collateral arteries. This can result in claudication, tissue damage, and eventually amputation. Conversely, healthy individuals undergo the growth and expansion of collateral arteries, or arteriogenesis, in order to compensate for reduced blood flow. Previous work from our laboratory has indicated that tumor endothelial marker 8 (TEM8) is necessary for arteriogenesis to occur in a mouse model of PAD. However, the mechanism behind this requirement is unknown. To address this gap in knowledge, we cloned the full length coding sequence (CDS) of the human TEM8 gene. The coding sequences were amplified in two parts via RT-PCR using RNA from HDMEC and T4 CHO cells. The amplified products were then ligated into the pCR4-TOPO TA vector. Sequence analysis verified the cloned products as human TEM8. Finally, the two cloned parts of human TEM8 were ligated together to create a full length human TEM8 clone. Introduce mutations to TEM8 CDS and analyze the effects of the mutations on potential relevant functions, such as adhesion, migration, polarization under fluid shear stress, and proliferation.

Funder Acknowledgement(s): This research opportunity was supported by Van Andel Research Institute, under Dr. Nick Duesbery and Dr. Jenn White, and the United Negro College Fund.

Faculty Advisor: Nick Duesbery,

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This material is based upon work supported by the National Science Foundation (NSF) under Grant No. DUE-1930047. Any opinions, findings, interpretations, conclusions or recommendations expressed in this material are those of its authors and do not represent the views of the AAAS Board of Directors, the Council of AAAS, AAAS’ membership or the National Science Foundation.

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