Biotechnology Commons^™

United States Regulation Of Stem Cell Research: Recasting Government's Role And Questions To Be Resolved, Owen C. B. Hughes, Alan L. Jakimo, Michael J. Malinowski

Journal Articles

This article directly addresses the stem cell controversy, but also the broader history and norms regarding the roles of federal and state government in U.S. science research funding.

Mapping The Functional Interaction Of Sco1 And Cox2 In Cytochrome Oxidase Biogenesis, Kevin Rigby, Paul A. Cobine, Oleh Khalimonchuk, Dennis R. Winge

Department of Biochemistry: Faculty Publications

Sco1 is implicated in the copper metallation of the Cu_A site in

Cox2 of cytochrome oxidase. The structure of Sco1 in the metallated

and apo-conformers revealed structural dynamics primarily

in an exposed region designated loop 8. The structural

dynamics of loop 8 in Sco1 suggests it may be an interface for

interactions with Cox17, the Cu(I) donor and/or Cox2. A series

of conserved residues in the sequence motif ²¹⁷KKYRVYF²²³ on

the leading edge of this loop are shown presently to be important

for yeast Sco1 function. Cells harboring Y219D, R220D, V221D,

and Y222D mutant Sco1 proteins failed …

Function And Redox State Of Mitochondrial Localized Cysteine-Rich Proteins Important In The Assembly Of Cytochrome C Oxidase, Oleh Khalimonchuk, Dennis R. Winge

Department of Biochemistry: Faculty Publications

The cytochrome c oxidase (CcO) complex of the mitochondrial respiratory chain exists within the mitochondrial inner membrane (IM). The biogenesis of the complex is a multi-faceted process requiring multiple assembly factors that function on both faces of the IM. Formation of the two copper centers of CcO occurs within the intermembrane space (IMS) and is dependent on assembly factors with critical cysteinyl thiolates. Two classes of assembly factors exist, one group being soluble IMS proteins and the second class being proteins tethered to the IM. A common motif in the soluble assembly factors is a duplicated Cx₉C sequence …

Pet191 Is A Cytochrome C Oxidase Assembly Factor In Saccharomyces Cerevisiae, Oleh Khalimonchuk, Kevin Rigby, Megan Bestwick, Fabien Pierrel, Paul A. Cobine, Dennis R. Winge

Department of Biochemistry: Faculty Publications

The twin-Cx₉C motif protein Pet191 is essential for cytochrome c oxidase maturation. The motif Cys residues are functionally important and appear to be present in disulfide linkages within a large oligomeric complex associated with the mitochondrial inner membrane. The import of Pet191 differs from that of other twin-Cx₉C motif class of proteins in being independent of the Mia40 pathway.

Coa2 Is An Assembly Factor For Yeast Cytochrome C Oxidase Biogenesis That Facilitates The Maturation Of Cox1, Fabien Pierrel, Oleh Khalimonchuk, Paul A. Cobine, Megan Bestwick, Dennis R. Winge

Department of Biochemistry: Faculty Publications

The assembly of cytochrome c oxidase (CcO) in yeast mitochondria is dependent on a new assembly factor designated Coa2. Coa2 was identified from its ability to suppress the respiratory deficiency of coa1 Δ and shy1 Δ cells. Coa1 and Shy1 function at an early step in maturation of the Cox1 subunit of CcO. Coa2 functions downstream of the Mss51-Coa1 step in Cox1 maturation and likely concurrent with the Shy1-related heme a₃ insertion into Cox1. Coa2 interacts with Shy1. Cells lacking Coa2 show a rapid degradation of newly synthesized Cox1. Rapid Cox1 proteolysis also occurs in shy1 Δ cells, suggesting …

Purification From Human Milk Of Matriptase Complexes With Secreted Serpins: Mechanism For Inhibition Of Matriptase Other Than Hai-1, I-Chu Tseng, Feng-Pai Chou, Sheng-Feng Su, Michael Oberst, Nandakumar Madayiputhiya, Ming-Shyue Lee, Jehng-Kang Wang, David E. Sloane, Michael Johnson, Chen-Yong Lin

Department of Biochemistry: Faculty Publications

Matriptase, a type 2 transmembrane serine protease, is predominately expressed by epithelial and carcinoma cells in which hepatocyte growth factor activator inhibitor 1 (HAI-1), a membrane-bound, Kunitz-type serine protease inhibitor, is also expressed. HAI-1 plays dual roles in the regulation of matriptase, as a conventional protease inhibitor and as a factor required for zymogen activation of matriptase. As a consequence, activation of matriptase is immediately followed by HAI-1-mediated inhibition, with the activated matriptase being sequestered into HAI-1 complexes. Matriptase is also expressed by peripheral blood leukocytes, such as monocytes and macrophages; however, in contrast to epithelial cells, monocytes and macrophages …

N-Glycan Modification In Aspergillus Species, Elke Kainz, Andreas Gallmetzer, Christian Hatzl, Juergen H. Nett, Huijuan Li, Thorsten Schinko, Robert Pachlinger, Harald Berger, Yazmid Reyes-Dominguez, Andreas Bernreiter, Tillmann Gerngross, Stefan Wildt, Joseph Strauss

Dartmouth Scholarship

The production by filamentous fungi of therapeutic glycoproteins intended for use in mammals is held back by the inherent difference in protein N-glycosylation and by the inability of the fungal cell to modify proteins with mammalian glycosylation structures. Here, we report protein N-glycan engineering in two Aspergillus species. We functionally expressed in the fungal hosts heterologous chimeric fusion proteins containing different localization peptides and catalytic domains. . This strategy allowed the isolation of a strain with a functional -1,2-mannosidase producing increased amounts of N-glycans of the Man 5 GlcNAc 2 type. This strain was further engineered by the introduction of …

High-Throughput Screening For Fatty Acid Uptake Inhibitors In Humanized Yeast Identifies Atypical Antipsychotic Drugs That Cause Dyslipidemias, Hong Li, Paul N. Black, Aalap Chokshi, Angel Sandoval-Alvarez, Ravi Vatsyayan, Whitney Sealls, Concetta Dirusso

Department of Biochemistry: Faculty Publications

Fatty acids are implicated in the development of dyslipidemias, leading to type 2 diabetes and cardiovascular disease. We used a standardized small compound library to screen humanized yeast to identify compounds that inhibit fatty acid transport protein (FATP)-mediated fatty acid uptake into cells. This screening procedure used live yeast cells expressing human FATP2 to identify small compounds that reduced the import of a fluorescent fatty acid analog, 4,4-difluoro-5-methyl-4-bora-3a,4a-diaza-s-indacene-3-dodecanoic acid (C1-BODIPY-C12). The library used consisted of 2,080 compounds with known biological activities. Of these, ~1.8% reduced cell-associated C1-BODIPY-C12 fluorescence and were selected as potential inhibitors of human FATP2- mediated fatty acid …

Comparative Functional Analysis Of Aquaporins/Glyceroporins In Mammals And Anurans, Carissa M. Krane, David L. Goldstein

Biology Faculty Publications

Maintenance of fluid homeostasis is critical to establishing and maintaining normal physiology. The landmark discovery of membrane water channels (aquaporins; AQPs) ushered in a new area in osmoregulatory biology that has drawn from and contributed to diverse branches of biology, from molecular biology and genomics to systems biology and evolution, and from microbial and plant biology to animal and translational physiology. As a result, the study of AQPs provides a unique and integrated backdrop for exploring the relationships between genes and genome systems, the regulation of gene expression, and the physiologic consequences of genetic variation. The wide species distribution of …

Water. New Waters And New Life, Juan Enriquez

New England Journal of Public Policy

An excerpt from an article about life sciences taken from the proceedings of the Education for Public Inquiry and International Citizenship (EPIIC) Symposium held at Tufts University in Massachusetts in February 2005 is presented.

Optimization Of Cutaneous Electrically Mediated Plasmid Dna Delivery Using Novel Electrode, L. C. Heller, M. J. Jaroszeski, D. Coppola, A. N. Mccray, J. Hickey, R. Heller

Bioelectrics Publications

The easy accessibility of skin makes it an excellent target for gene transfer protocols. To take advantage of skin as a target for gene transfer, it is important to establish an efficient and reproducible delivery system. Electroporation is an established technique for enhancing plasmid delivery to many tissues in vivo. A critical component of this technique is the electrode configuration. Electroporation parameters were optimized for transgene expression with minimal tissue damage with a novel electrode. The highest transgene expression and efficiency of individual cell transformation with minimal damage was produced with eight 150 ms pulses at field strength of …

Human Genome, South Dakota State University

Biotechnology

Bibliography and photograph of a display of government documents from South Dakota State University.

Integration Of Gis And Bentgrass Ecology For Ecological Risk Assessment., Carol Auer, Collin Ahrens

Plant Science Presentations and Proceedings

All new technologies carry both benefits and risks. Genetically-modified plants must be assessed before they are released into the environment. Our research examines plant gene flow and the potential impacts from environmental release of genetically-modified plants. Plant gene flow is a natural process that occurs when pollen from one plant lands on the flower of another plant and produces a hybrid offspring. Gene flow can produce hybrid offspring with new traits that could change the ability of the plant to survive and spread. If hybrid offspring have some advantage in the environment, they could become invasive and/or affect other components …

Selective Cholesterol Dynamics Between Lipoproteins And Caveolae/Lipid Rafts, Stephen M. Storey, Adalberto M. Gallegos, Barbara P. Atshaves, Avery L. Mcintosh, Gregory G. Martin, Rebecca D. Parr, Kerstin K. Landrock, Ann B. Kier, Judith M. Ball, Friedhelm Schroeder

Faculty Publications

Although low-density lipoprotein (LDL) receptor-mediated cholesterol uptake through clathrin-coated pits is now well understood, the molecular details and organizing principles for selective cholesterol uptake/efflux (reverse cholesterol transport, RCT) from peripheral cells remain to be resolved. It is not yet completely clear whether RCT between serum lipoproteins and the plasma membrane occurs primarily through lipid rafts/caveolae or from non-raft domains. To begin to address these issues, lipid raft/caveolae-, caveolae-, and non-raft-enriched fractions were resolved from purified plasma membranes isolated from L-cell fibroblasts and MDCK cells by detergent-free affinity chromatography and compared with detergent-resistant membranes isolated from the same cells. Fluorescent sterol …

Over-Expression Of Udp-Glucose Pyrophosphorylase In Hybrid Poplar Affects Carbon Allocation, Heather Coleman, Thomas Canam, Kyu-Young Kang, David Ellis, Shawn Mansfield

Faculty Research & Creative Activity

The effects of the over-expression of the Acetobacter xylinum UDP-glucose pyrophosphorylase (UGPase) under the control of the tandem repeat Cauliflower Mosaic Virus promoter (2335S) on plant metabolism and growth were investigated in hybrid poplar (Populus alba3grandidentata). Transcript levels, enzyme activity, growth parameters, leaf morphology, structural and soluble carbohydrates, and soluble metabolite levels were quantified in both transgenic and wild-type trees. Transgenic 2335S::UGPase poplar showed impaired growth rates, displaying reduced height growth and stem diameter. Morphologically, 2335S::UGPase trees had elongated axial shoots, and leaves that were substantially smaller in size when compared with wild-type trees at equivalent developmental stages. Biochemical analysis …

Benjamin In Paris: Weak Messianism And Memories Of The Oppressed, Jules Simon

Selected Works Temporary Series

No abstract provided.

Full-Length, Glycosylated Nsp4 Is Localized To Plasma Membrane Caveolae By A Novel Raft Isolation Technique, Stephen M. Storey, Thomas F. Gibbons, Cecelia V. Williams, Rebecca D. Parr, Friedhelm Schroeder, Judith M. Ball

Faculty Publications

Rotavirus NSP4, initially characterized as an endoplasmic reticulum intracellular receptor, is a multifunctional viral enterotoxin that induces diarrhea in murine pups. There have been recent reports of the secretion of a cleaved NSP4 fragment (residues 112 to 175) and of the association of NSP4 with LC3-positive autophagosomes, raft membranes, and microtubules. To determine if NSP4 traffics to a specific subset of rafts at the plasma membrane, we isolated caveolae from plasma membrane-enriched material that yielded caveola membranes free of endoplasmic reticulum and nonraft plasma membrane markers. Analyses of the newly isolated caveolae from rotavirus-infected MDCK cells revealed full-length, high-mannose glycosylated …

Over-Expression Of Udp-Glucose Pyrophosphorylase In Hybrid Poplar Affects Carbon Allocation, Heather D. Coleman, Thomas Canam, Kyu-Young Kang, David D. Ellis, Shawn D. Mansfield

Faculty Research & Creative Activity

The effects of the over-expression of the Acetobacter xylinum UDP-glucose pyrophosphorylase (UGPase) under the control of the tandem repeat Cauliflower Mosaic Virus promoter (2335S) on plant metabolism and growth were investigated in hybrid poplar (Populus alba3grandidentata). Transcript levels, enzyme activity, growth parameters, leaf morphology, structural and soluble carbohydrates, and soluble metabolite levels were quantified in both transgenic and wild-type trees. Transgenic 2335S::UGPase poplar showed impaired growth rates, displaying reduced height growth and stem diameter. Morphologically, 2335S::UGPase trees had elongated axial shoots, and leaves that were substantially smaller in size when compared with wild-type trees at equivalent developmental stages. Biochemical analysis …

Biotechnology: The Genetics Revolution, Minnesota State University, Mankato

Biotechnology

Bibliography and photographs of a display of government documents from Minnesota State University, Mankato.

Synthetic Biology: Caught Between Property Rights, The Public Domain, And The Commons, Arti K. Rai, James Boyle

Faculty Scholarship

Synthetic biologists aim to make biology a true engineering discipline. In the same way that electrical engineers rely on standard capacitors and resistors, or computer programmers rely on modular blocks of code, synthetic biologists wish to create an array of modular biological parts that can be readily synthesized and mixed together in different combinations. Synthetic biology has already produced important results, including more accurate AIDS tests and the possibility of unlimited supplies of previously scarce drugs for malaria. Proponents hope to use synthetic organisms to produce not only medically relevant chemicals but also a large variety of industrial materials, including …

Analysis Of Optimum Location, Viability, And Potential Effects Of Wind Farm Construction In Maine, Elizabeth Benson, Ryan Scott

Undergraduate Research Symposium (UGRS)

Maine has the highest potential for wind energy in New England and falls within the top twenty states in the nation. It falls just behind Wisconsin and California with an estimate electrical output of 56 billion kWhs. The geological makeup of Maine’s mountains in the western part of the state, and the exposed coastline provide opportune areas to capture wind and convert it into energy. The information included in this poster will suggest the most likely areas for wind development based on a number of factors as recommended by the American Wind Energy Association.

Potential For Stimulating Host Anti-Tumor Immune Response Via Rnai-Mediated Local Foxp3 Knockdown, N. Klaiber

Chemistry & Biochemistry Faculty Publications

Neoplastic growths represent a unique challenge for the host immune system. As they are indeed derived from self, many of the same mechanisms operating to prevent autoimmunity also provide an umbrella beneath which malignant cells are free to proliferate.1 Central among these immune regulatory boundaries are an influential subset of lymphocytes known as T regs. Hypothesized to exist decades ago, yet eluding definitive characterization until relatively recently, T regs have been demonstrated to play a crucial role in the proper functioning of the immune system as a whole. They may also, however, represent one of the primary obstacles to successful …

Sensory Genes And Mate Choice: Evidence That Duplications, Mutations, And Adaptive Evolution Alter Variation In Mating Cue Genes And Their Receptors, Lisa Horth

Biological Sciences Faculty Publications

Fascinating new data, revealed through gene sequencing, comparative genomics, and genetic engineering, precisely establish which genes are involved in mate choice and mating activity—behaviors that are surprisingly understudied from a genetic perspective. Discussed here are some of the recently identified visual and chemosensory genes that are involved in mate choice and mating behavior. These genes’ products are involved in the production, transmission, and receipt of crucial sensory mate-choice cues that affect fitness. This review exposes newfound evidence that alternative splicing, gene-expression pattern changes, and molecular genetic variation in sensory genes are crucial for both intra- and interspecific mate choice and …

Biotechnology, Life Sciences And Control Engineering:Some Synthesis Issues, Aidan O'Dwyer

Conference papers

The synthesis of biotechnology, life sciences and control engineering is receiving increasing interest. The purpose of this contribution (and the associated poster) is to raise awareness of some of this synthesis work.

Assessing The Universal Declaration On Bioethics And Human Rights: Implications For Human Dignity And The Respect For Human Life, O. Carter Snead

Journal Articles

As advances in biomedical science and biotechnology continue to confer vast new powers to subdue nature for “the relief of man’s estate,” vexed moral and ethical questions inevitably arise about the use of such capacities. To what ends should they be applied? How might they be abused? Are they safe? What is the human meaning of such new powers? Might their use threaten human dignity? What does scientific freedom demand? What does justice require? Who should have access to the fruits of such scientific advances?

In democratic regimes, these questions do not long remain the subject of mere academic reflection …

A New Strategy For Assessing Selenoprotein Function: Sirna Knockdown/Knock-In Targeting The 3'-Utr, Min-Hyuk Yoo, Xue-Ming Xu, Anton A. Turanov, Bradley A. Carlson, Vadim N. Gladyshev, Dolph L. Hatfield

Department of Biochemistry: Faculty Publications

Selenocysteine insertion into protein in mammalian cells requires RNA elements in the 3'-untranslated regions (3'-UTRs) of selenoprotein genes. The occurrence of these conserved sequences should make selenoproteins particularly amenable for knockdown/knock-in strategies to examine selenoprotein functions. Herein, we utilized the 3'-UTR of various selenoproteins to knock down their expression using siRNAs and then knock in expression using constructs containing mutations within the target region. Thioredoxin reductase 1 (TR1) knockdown in a mouse kidney cell line resulted in the cells growing about 10% more slowly, being more sensitive to UV radiation, and having increased apoptosis in response to UV than control …

Mechanistic Studies Of The Long Chain Acyl-Coa Synthetase Faa1p From Saccharomyces Cerevisiae, Hong Li, Elaina M. Melton, Steven Quackenbush, Concetta C. Dirusso, Paul N. Black

Department of Biochemistry: Faculty Publications

Long chain acyl-CoA synthetase (ACSL; fatty acid CoA ligase: AMP forming; EC 6.2.1.3) catalyzes the formation of acyl-CoA through a process, which requires fatty acid, ATP and coenzymeA as substrates. In the yeast Saccharomyces cerevisiae the principal ACSL is Faa1p (encoded by the FAA1 gene). The preferred substrates for this enzyme are cis-monounsaturated long chain fatty acids. Our previous work has shown Faa1p is a principal component of a fatty acid transport/activation complex that also includes the fatty acid transport protein Fat1p. In the present work hexameric histidine tagged Faa1p was purified to homogeneity through a two-step process in the …

Evidence For A Pro-Oxidant Intermediate In The Assembly Of Cytochrome Oxidase, Oleh Khalimonchuk, Amanda Bird, Dennis R. Winge

Department of Biochemistry: Faculty Publications

The hydrogen peroxide sensitivity of cells lacking two proteins,

Sco1 and Cox11, important in the assembly of cytochrome

c oxidase (CcO), is shown to arise from the transient accumulation

of a pro-oxidant heme A-Cox1 stalled intermediate. The

peroxide sensitivity of these cells is abrogated by a reduction in

either Cox1 expression or hemeAformation but exacerbated by

either enhanced Cox1 expression or heme A production arising

from overexpression of COX15. Sco1 and Cox11 are implicated

in the formation of the Cu_A and Cu_B sites of CcO, respectively.

The respective wild-type genes suppress the peroxide sensitivities

of sco1∆ …

Coa1 Links The Mss51 Post-Translational Function To Cox1 Cofactor Insertion In Cytochrome C Oxidase Assembly, Fabien Pierrel, Megan L. Bestwick, Paul A. Cobine, Oleh Khalimonchuk, Julia A. Cricco, Dennis R. Winge

Department of Biochemistry: Faculty Publications

The assembly of cytochrome c oxidase (CcO) in yeast mitochondria is shown to be dependent on a new assembly factor designated Coa1 that associates with the mitochondrial inner membrane. Translation of the mitochondrial-encoded subunits of CcO occurs normally in coa1Δ cells, but these subunits fail to accumulate. The respiratory defect in coa1Δ cells is suppressed by high-copy MSS51, MDJ1 and COX10. Mss51 functions in Cox1 translation and elongation, whereas Cox10 participates in the biosynthesis of heme a, a key cofactor of CcO. Respiration in coa1Δ and shy1Δ cells is enhanced when Mss51 and …

Synthetic Biology: The Intellectual Property Puzzle, Arti K. Rai, Sapna Kumar

Faculty Scholarship

Synthetic biology, which operates at the intersection of biotechnology and information technology, has the potential to raise, in a particularly acute manner, the intellectual property problems that exist in both fields. A preliminary patent landscape reveals problematic foundational patents that could, if licensed and enforced inappropriately, impede the potential of the technology. The landscape also reveals a proliferation of patents on basic synthetic biology "parts" that could create transaction cost heavy patent thickets. Both foundational patents and patent thickets are likely to be particularly problematic to the extent they read on standards that synthetic biologists would like to establish. Synthetic …