Biotechnology Commons^™

Analysis Of Leigh Syndrome Mutations In The Yeast Surf1 Homolog Reveals A New Member Of The Cytochrome Oxidase Assembly Factor Family, Megan Bestwick, Mi-Young Jeong, Oleh Khalimonchuk, Hyung Kim, Dennis R. Winge

Department of Biochemistry: Faculty Publications

Three missense SURF1 mutations identified in patients with Leigh syndrome (LS) were evaluated in the yeast homolog Shy1 protein. Introduction of two of the Leigh mutations, F²⁴⁹T and Y³⁴⁴D, in Shy1 failed to significantly attenuate the function of Shy1 in cytochrome c oxidase (CcO) biogenesis as seen with the human mutations. In contrast, a G¹³⁷E substitution in Shy1 results in a nonfunctional protein conferring a CcO deficiency. The G¹³⁷E Shy1 mutant phenocopied shy1 Δ cells in impaired Cox1 hemylation and low mitochondrial copper. A genetic screen for allele-specific suppressors of the G137E …

The Role Of Coa2 In Hemylation Of Yeast Cox1 Revealed By Its Genetic Interaction With Cox10, Megan Bestwick, Oleh Khalimonchuk, Fabien Pierrel, Dennis R. Winge

Department of Biochemistry: Faculty Publications

Saccharomyces cerevisiae cells lacking the cytochrome c oxidase (CcO) assembly factor Coa2 are impaired in Cox1 maturation and exhibit a rapid degradation of newly synthesized Cox1. The respiratory deficiency of coa2 Δ cells is suppressed either by the presence of a mutant allele of the Cox10 farnesyl transferase involved in heme a biosynthesis or through impaired proteolysis by the disruption of the mitochondrial Oma1 protease. Cox10 with an N196K substitution functions as a robust gain-of-function suppressor of the respiratory deficiency of coa2 Δ cells but lacks suppressor activity for two other CcO assembly mutant strains, the coa1 Δ and shy1 …

Transgenic Rat Model Of Neurodegeneration Caused By Mutation In The Tdp Gene, Hongxia Zhou, Cao Huang, Han Chen, Dian Wang, Carlisle P. Landel, Pedro Yuxing Xia, Robert Bowser, Yong-Jian Liu, Xu Gang Xia

Nebraska Center for Biotechnology: Faculty and Staff Publications

TDP-43 proteinopathies have been observed in a wide range of neurodegenerative diseases. Mutations in the gene encoding TDP-43 (i.e., TDP) have been identified in amyotrophic lateral sclerosis (ALS) and in frontotemporal lobe degeneration associated with motor neuron disease. To study the consequences of TDP mutation in an intact system, we created transgenic rats expressing normal human TDP or a mutant form of human TDP with a M337V substitution. Overexpression of mutant, but not normal, TDP caused widespread neurodegeneration that predominantly affected the motor system. TDP mutation reproduced ALS phenotypes in transgenic rats, as seen by progressive degeneration of motor neurons …

Biosensors For Biodiesel Quality Sensing, M. Z. Hossain, D. S. Shrestha, Maurice G. Kleve

Journal of the Arkansas Academy of Science

A biosensor is an analytical device that uses biomaterials as elements of the sensing system and converts a biological response into an electrical signal. Biodiesel is a bio-based alternative, biodegradable, renewable, nontoxic diesel fuel made from a chemical reaction between alcohol (usually methanol or ethanol) and plant oil or animal fat. A need to provide accurate, real-time information for the quality sensing of biodiesel properties such as free and total glycerol has led to an ever-increasing demand for biosensor development. Being able to monitor specific physical and chemical properties is the prerequisite for developing a biosensor for quality sensing of …

Plasmid Injection And Application Of Electric Pulses Alter Endogenous Mrna And Protein Expression In B16.F10 Mouse Melanomas, L. C. Heller, Y. L. Cruz, B. Ferraro, H. Yang, R. Heller

Bioelectrics Publications

The application of electric pulses to tissues causes cell membrane destabilization, allowing exogenous molecules to enter the cells. This delivery technique can be used for plasmid gene therapy. Reporter gene expression after plasmid delivery with eight representative published protocols was compared in B16.F10 mouse melanoma tumors. This expression varied significantly based on the pulse parameters utilized for delivery. To observe the possible influence of plasmid injection and/or pulse application on endogenous gene expression, levels of stress-related mRNAs 4 and 24 h after delivery were determined by PCR array. Increases in mRNA levels for several inflammatory chemokines and cytokines were observed …

Electroporation-Mediated Delivery Of A Naked Dna Plasmid Expressing Vegf To The Porcine Heart Enhances Protein Expression, W. G. Marshall Jr., B. A. Boone, J. D. Burgos, S. I. Gografe, M. K. Baldwin, M. L. Danielson, M. J. Larson, D. R. Caretto, Y. Cruz, B. Ferraro, L. C. Heller, K. E. Ugen, M. J. Jaroszeski, R. Heller

Bioelectrics Publications

Gene therapy is an attractive method for the treatment of cardiovascular disease. However, using current strategies, induction of gene expression at therapeutic levels is often inefficient. In this study, we show a novel electroporation (EP) method to enhance the delivery of a plasmid expressing an angiogenic growth factor (vascular endothelial growth factor, VEGF), which is a molecule previously documented to stimulate revascularization in coronary artery disease. DNA expression plasmids were delivered in vivo to the porcine heart with or without coadministered EP to determine the potential effect of electrically mediated delivery. The results showed that plasmid delivery through EP significantly …

Mzm1 Influences A Labile Pool Of Mitochondrial Zinc Important For Respiratory Function, Aaron Atkinson, Oleh Khalimonchuk, Pamela Smith, Hana Sabic, David Eide, Dennis R. Winge

Department of Biochemistry: Faculty Publications

Zinc is essential for function of mitochondria as a cofactor for

several matrix zinc metalloproteins. We demonstrate that a

labile cationic zinc component of low molecular mass exists in

the yeast mitochondrial matrix. This zinc pool is homeostatically

regulated in response to the cellular zinc status. This pool

of zinc is functionally important because matrix targeting of a

cytosolic zinc-binding protein reduces the level of labile zinc

and interferes with mitochondrial respiratory function. We

identified a series of proteins that modulate the matrix zinc

pool, one of which is a novel conserved mitochondrial protein

designated Mzm1. Mutant mzm1∆ cells …

Purification And Analysis Of Bacterial-And Plant-Derived Scfv Antibodies For The Detection Of Cucumber Mosaic Virus, Wong C.F.Y.

Student Works (2010-2019)

abstract N/A

Antibactertial Property And Toxcity Study Of Medicinal Plant Tinospora Crispa L., Altammar Khadijah Ahmed Y.

Student Works (2010-2019)

Tinospora crispa is a medically important herb used for centuries in traditional medicine for the treatment of various ailments. The aim of this study was to investigate the safety of the aqueous and alcoholic extract of Tinospora crispa by determining its toxicological effects after acute administration in rats, and to investigate the antibacterial activity of both extracts. Disc diffusion and broth dilution methods were carried out in determining the antibacterial effects of the plant extract on the pure cultures of clinical isolates of 8 MRSA and S. aureus ATCC strain. The disc-diffusion method of the ethanolic extracts showed inhibition zone …

Biolistic Transformation Of Oil Palm Using The Phosphomannose Isomerase (Pmi) Gene As The Positive Selectable Marker., Bahariah Bohari

Student Works (2010-2019)

The selectable marker system based on the Escherichia coli phosphomannose isomerase (pmi) gene was adapted for genetic transformation of oil palm. This system makes use of the pmi gene that encodes phosphomannose isomerase, which converts mannose-6-phosphate to fructose-6-phosphate and uses mannose as the selection agent. This is to anticipate the future requirement of using non-antibiotic resistance genes for the commercialization of transgenic oil palm. The use of antibiotic or herbicide-based selection systems has caused much public concern due to inadequate knowledge of the agents’ impact on the environment and on human health. In this study, four transformation vectors, namely pMI3, …

Studies On The Bioactive Properties Of Selected Plant And Fermented Extracts., Saravana Kumar Sinniah

Student Works (2010-2019)

The aim of this study was to isolate protein/peptide and secondary metabolites from plant and fermented sources. The extracts were used to test for antimicrobial and antioxidant properties. In the current study protein/peptide extracts of Allium sativum showed promising results with a 15mm inhibition zone against Escherichia coli, 28mm against Staphylococcus aureus, 16.3mm against Bacillus cereus and 9mm against Pseudomonas aeruginosa. The minimum bacteriocidal concentration (MBC) results showed that Allium sativum protein/peptide extract is only bacteriostatic and not bacteriocidal. Andrographis paniculata showed the most promising result for the ethanolic extracts with an inhibition zone of 14mm against B. cereus, 11.5mm …

Estimation Of The Product Yield And Collection Efficiency Of Peripheral Blood Stem Cell Harvesting Using Peripheral Blood Cd34[+] Cells Quantification., Habsah Aziz

Student Works (2010-2019)

Peripheral blood stem cells (PBSC) have been used for transplantation since the early 1990s. CD34+ has been used as a marker of hematopoietic stem cells to determine the optimal timing for PBSC harvesting. However, a wide ranges of CD34+ cells cut-off values from 8/ul to 66/ul have been used to initiate PBSC harvesting in autologous patients. We performed here a study to determine the peripheral blood CD34+ cells cut-off value for the prediction of stem cell yield of 2 x106 CD34+ cells/kg body weight patients, to study the correlation between peripheral blood CD34+ cells and stem cell yield, to study …

Diversity And Antioxidant Activity Of Trametes Fr. In Malaysia., Noraswati Mohd Nor Rashid

Student Works (2010-2019)

Research on antioxidant properties as a natural antioxidant, particularly in polypores, have been widely done since the past decade. It is known that antioxidant capacity is essential in protecting organisms and cells from damage brought on by oxidative which is believed to be the cause for ageing and degenerative diseases. In this study, biodiversity and antioxidant activity of Malaysian Trametes spp. which is from the group of polypores were investigated. Polypores were collected in Peninsular Malaysia forests from the year 2006 until 2007. The species were identified by taxonomic keys based on macromorphology and micromorphology analysis. Trametes hirsuta, T. pocas, …

Cytotoxic And Anti-Human Papillomavirus Activites In Selected Vegetables., Maizatul Azmah Ab. Latiff

Student Works (2010-2019)

An appropriate way to control cancer is chemoprevention as a large number of compounds have been observed to prevent the occurrence of cancer and variety of mechanisms exists for producing such protective effects. Numerous epidemiological studies have shown that diets rich in vegetables are associated with a decreased risk of cancer. Chemoprevention involves the use of natural or laboratory made substances to prevent cancer or reduce risk cancer.The E6 oncoprotein from oncogenic Human papillomaviruses (HPVs) has exhibit functions in tumorigenesis, regulation of transcriptions and apoptosis. E6 product from oncogenic viral type like HPV 16 can bind the tumor suppressor p53 …

Synthesis, Characterization And Biological Activities Of Three Indole Schiff Bases And Their Copper And Nickel Complexes., Nazzatush Shimar Jamaludin

Student Works (2010-2019)

Three derivatives of indole-3-carboxaldehyde with indole-3-acetic hydrazide and their copper(II) and nickel(II) complexes were synthesized. The Schiff base compounds were varied on the substituent where each compound holds a hydrogen, chlorine and bromine atom on the 5th position of indole-3-carboxaldehyde. The structures of the ligands and their complexes were elucidated by various characterization methods. All compounds were screened for their biological effects on neuroprotective screening (in vitro), acute toxicity test (in vivo), ethanol-induced gastric ulcer (anti-ulcer) and glucose tolerance test (antidiabetic). All compounds except bromine-containing compounds were not dose dependent while compounds with bromine displayed significant results, acting as inhibitory …

In Vitro Cultures For Production Of Zerumbone And Mass Propagation In Zingiber Zerumbet Smith., Farah Diana Idris

Student Works (2010-2019)

Lempoyang (Zingiber zerumbet Smith.) is an important plant which has been propagated because of its medicinal values. Lempoyang is widely used as a folk medicine in Asian countries such as Malaysia, Indonesia, India and Taiwan. Zerumbone, a cyclic sesqueterpene found to be the major and main bioactive compound that gives great potential in medicinal purposes especially in the treatment of chronic diseases based on recent reports. Recently, it was found that zerumbone inhibited the growth of human leukemia cell line, HL-60 cell. Even though there are so many great values contain in the lempoyang but it is hard to obtain …

Agrobacterium-Mediated Transformation Of Cryptocoryne Willisii With Gus And Mgfp 5 Reporter Genes / Wong Sher Ming, Wong Sher Ming

Student Works (2010-2019)

In this study, a simple method using Murashige and Skoog basal medium (1962) supplemented with 6-BA alone was developed for direct multiple shoots induction from shoot base culture of C. willisii. Shoots proliferation and elongation were achieved on a single medium without the need of subculturing. The optimum concentration of 6-BA for direct shoots regeneration of C. willisii was investigated. It was found to be 6 mg/ L. Optimum media was then used for regenerate and propagate of transformed C. willisii. On the other hand, ransformation of C. willisii with Agrobacterium tumefaciens strain LBA4404 harbouring pCAMBIA1304 was performed. Effects on …

A Study On Cell Suspension Cultures For Micropropagation And The Production Of Carpaine From Papaya (Carica Papaya L. Var. Eksotika)., Mohamad Fhaizal Mohamad Bukhori

Student Works (2010-2019)

Carica papaya L. is a member of Caricaceae family with four native genera. This fruit is economically important as in multi-purpose fruit produced for dessert and vegetable and is highly in demand for pharmaconomical trade particularly the plant compound as source of medicine. This species is a popular fruit crop in the tropical and sub-tropical regions of Malaysia, Thailand, Indonesia and the Philippines. It is also one of the most widely grown fruits in Malaysia and is mainly cultivated in Johor, Perak and Selangor. The Eksotika papaya variety was introduced to the farmers in the late eighties and this resulted …

Antibactertial Property And Toxcity Study Of Medicinal Plant Tinospora Crispa L., Altammar Khadijah Ahmed Y.

Student Works (2010-2019)

Tinospora crispa is a medically important herb used for centuries in traditional medicine for the treatment of various ailments. The aim of this study was to investigate the safety of the aqueous and alcoholic extract of Tinospora crispa by determining its toxicological effects after acute administration in rats, and to investigate the antibacterial activity of both extracts. Disc diffusion and broth dilution methods were carried out in determining the antibacterial effects of the plant extract on the pure cultures of clinical isolates of 8 MRSA and S. aureus ATCC strain. The disc-diffusion method of the ethanolic extracts showed inhibition zone …

Evolution Of New Enzymatic Function By Structural Modulation Of Cysteine Reactivity In Pseudomonas Fluorescens Isocyanide Hydratase, Mahadevan Lakshminarasimhan, Peter Madzelan, Ruth Nan, Nicole Marie Milkovic, Mark A. Wilson

Department of Biochemistry: Faculty Publications

Isocyanide (formerly isonitrile) hydratase (EC 4.2.1.103) is an enzyme of the DJ-1 superfamily that hydrates isocyanides to yield the corresponding N-formamide. In order to understand the structural basis for isocyanide hydratase (ICH) catalysis, we determined the crystal structures of wild-type and several site-directed mutants of Pseudomonas fluorescens ICH at resolutions ranging from 1.0 to 1.9 Å. We also developed a simple UV-visible spectrophotometric assay for ICH activity using 2-naphthyl isocyanide as a substrate. ICH contains a highly conserved cysteine residue (Cys101) that is required for catalysis and interacts with Asp17, Thr102, and an ordered water molecule in the active …