http://espace.library.uq.edu.au/ The University of Queensland en Fez http://blogs.law.harvard.edu/tech/rss http://espace.library.uq.edu.au/view/UQ:281146 2012-09-04T17:18:45Z Li, Feng og Clegg, Jack K. og Goux-Capes, Laurence og Chastanet, Guillaume og D'Alessandro, Deanna M. og Letard, Jean-Francois og Kepert, Cameron J. http://espace.library.uq.edu.au/view/UQ:148494 2008-06-06T00:00:00Z Hwang, J. og Lee, H. og Maheshwari, H. og Smith, R. W. og Kroon, P. A. http://espace.library.uq.edu.au/view/UQ:112689 Burkholderia pseudomallei, the etiological agent of melioidosis, causes significant mortality in endemic regions, but little is known regarding the immune mechanisms required for successful protective immunity. To establish a model of immunization that could be used to study this we screened a library of B. pseudomallei strains for immunogenicity in mice. BALB/c mice were immunized with test strains, and 2 weeks later were given a lethal challenge (LC) of virulent B. pseudomallei. Among 49 strains tested, a single strain, CL04, exhibited strong immunoprotective capacity. Interestingly, CL04 had been cultured from a patient with chronic colonization of B. pseudomallei, which is a rare phenomenon. Mice immunized with 0.1 x LD50 (5 x 10(3) CFU) of CL04 had significantly better survival and lower bacterial loads after LC compared to naive controls. Dose-response analysis demonstrated more robust immunity after higher immunizing doses, and bacterial inactivation by gamma irradiation diminished the protective effect, indicating a requirement for viable organism for immunity. CL04-induced immunity was demonstrated both in B. pseudomallei-susceptible BALB/c and -resistant C57BL/6 mice. We investigated the gene profile of CL04-induced immunity by analyzing responses to immunization using cDNA microarray. Unique responses involving granulocyte macrophaloe colony stimulating factor (GM-CSF), the proapoptotic regulator Bad and cyclin-dependent kinase (CDK5) were detected in immunized mice, but these responses were absent in naive-LC mice. Further, responses differed between mouse strains, indicating dependence on host genetic background. This model will be useful in identifying elements of the immune response required for successful adaptive immunity against B. pseudomallei. (c) 2005 Elsevier SAS. All rights reserved. 2007-09-19T18:57:42Z Ulett, Glen C. og Labrooy, Justin T. og Currie, Bart J. og Barnes, Jodie L. og Ketheesan, Natkunam http://espace.library.uq.edu.au/view/UQ:191597 We have developed a three-dimensional model of the α1 homomeric glycine receptor by using Brownian dynamics simulations to account for its observed physiological properties. The model channel contains a large external vestibule and a shallow internal vestibule, connected by a narrow, cylindrical selectivity filter. Three rings of charged residues from the pore-lining M2 domain are modeled as point charges in the protein. Our simulations reproduce many of the key features of the channel, such as the current–voltage profiles, permeability ratios, and ion selectivity. When we replace the ring of alanine residues lining the selectivity filter with glutamates, the mutant model channel becomes permeable to cations, as observed experimentally. In this mutation, anions act as chaperones for sodium ions in the extracellular vestibule, and together they penetrate deep inside the channel against a steep energy barrier encountered by unaccompanied ions. Two subsequent amino acid mutations increase the cation permeability, enabling monovalent cations to permeate through the channel unaided and divalent cations to permeate when chaperoned by anions. These results illustrate the key structural features and underlying mechanism for charge selectivity in the glycine receptor. 2010-01-07T00:00:00Z O'Mara, Megan og Barry, Peter H. og Chung, Shin-Ho http://espace.library.uq.edu.au/view/UQ:269352 2012-03-08T00:00:00Z Thompson, M. F. og Mills, P. C. og Schembri, M. A. og Trott, D. J. http://espace.library.uq.edu.au/view/UQ:282607 2012-09-30T00:04:39Z Thompson, Mary F. og Schembri, Mark A. og Mills, Paul C. og Trott, Darren J. http://espace.library.uq.edu.au/view/UQ:112743 The formation, relative stability, and possible stoichiometries of two (self-) complementary peptide sequences (B and E) designed to form either a parallel homodimeric (B + B) or an antiparallel heterodimeric ( B + E) coiled coil have been investigated. Peptide B shows a characteristic coiled coil pattern in circular dichroism spectra at pH 7.4, whereas peptide E is apparently random coiled under these conditions. The peptides are complementary to each other, with peptide E forming a coiled coil when mixed with peptide B. Molecular dynamics simulations show that combinations of B + B and B + E readily form a dimeric coiled coil, whereas E + E does not fall in line with the experimental data. However, the simulations strongly suggest the preferred orientation of the helices in the homodimeric coiled coil is antiparallel, with interactions at the interface quite different to that of the idealized model. In addition, molecular dynamics simulations suggest equilibrium between dimers, trimers, and tetramers of alpha-helices for peptide B. 2007-09-19T00:00:00Z Pineiro, A. og Villa, A. og Vagt, T. og Koksch, B. og Mark, A. E. http://espace.library.uq.edu.au/view/UQ:111990 HIV-1 protease is most active under weakly acidic conditions (pH 3.5-6.5), when the catalytic Asp25 and Asp25' residues share 1 proton. At neutral pH, this proton is lost and the stability of the structure is reduced. Here we present an investigation of the effect of pH on the dynamics of HIV-1 protease using MD simulation techniques. MD simulations of the solvated HIV-1 protease with the Asp25/25' residues monoprotonated and deprotonated have been performed. In addition we investigated the effect of the inclusion of Na+ and Cl- ions to mimic physiological salt conditions. The simulations of the monoprotonated form and deprotonated form including Na+ show very similar behavior. In both cases the protein remained stable in the compact, "self-blocked" conformation in which the active site is blocked by the tips of the flaps. In the deprotonated system a Na+ ion binds tightly to the catalytic dyad shielding the repulsion between the COO- groups. Ab initio calculations also suggest the geometry of the active site with the Na+ bound closely resembles that of the monoprotonated case. In the simulations of the deprotonated form (without Na+ ions), a water molecule bound between the Asp25 Asp25' side-chains. This disrupted the dimerization interface and eventually led to a fully open conformation. (C) 2004 Wiley-Liss, Inc. 2007-09-19T00:00:00Z Kovalskyy, D. og Dubyna, V. og Mark, A. E. og Kornelyuk, A. http://espace.library.uq.edu.au/view/UQ:95354 2007-08-23T00:00:00Z Guyatt, K. J. og Twin, J. og Smith, G. og Smith, I. og Mackenzie, J. S. og Young, P. http://espace.library.uq.edu.au/view/UQ:65137 2007-08-14T19:38:37Z Guyatt, Kimberley J. og Twin, Jimmy og Davis, Patricia og Holmes, Edward C. og Smith, Greg A. og Smith, Ina L. og Mackenzie, John S. og Young, Peter L. http://espace.library.uq.edu.au/view/UQ:263567 2011-12-16T10:32:18Z McDevitt, Christopher A. og Ogunniyi, Abiodun D. og Valkov, Eugene og Lawrence, Michael C. og Kobe, Bostjan og McEwan, Alastair G. og Paton, James C. http://espace.library.uq.edu.au/view/UQ:97229 2007-08-24T00:00:00Z Smith, R. W. og Shan, J. og Munro, T. og Barbarese, E. og Carson, J. H. http://espace.library.uq.edu.au/view/UQ:58641 A new lepocreadiid genus, Amphicreadium, is erected for the species A. denspeniculus n. sp. from Acanthaluteres vittiger and for an unnamed species from Meuschenia freycineti, both from off northern Tasmania. The new genus is distinguished from all other members of its family by its amphistomatous body plan. 2007-08-14T00:00:00Z Bray, R. A. og Cribb, T. H. http://espace.library.uq.edu.au/view/UQ:286091 Locked nucleic acids (LNA) confer high thermal stability and nuclease resistance to oligonucleotides. The discovery of polymerases that accept LNA triphosphates has led us to propose a scheme for the amplification and re-generation of LNA-containing oligonucleotide libraries. Such libraries could be used for in vitro selection of e.g., native LNA aptamers. We maintained an oligonucleotide library encoding 40 randomized positions with LNA ATP, GTP, CTP, and TTP for 7 rounds of ‘mock’ in vitro selection in the absence of a target and analyzed the sequence composition after rounds 1, 4 and 7. We observed a decrease in LNA-A content from 20.5% in round 1 to 6.6% in round 7. This decrease was accompanied by a substantial bias against successive LNA-As (poly-LNA adenosine tracts) and a relative over-representation of single LNA-As. Maintaining a library with LNA TTP yielded similar results. Together, these results suggest that dispersed LNA monomers are tolerated in our in vitro selection protocol, and that LNA-modified libraries can be sustained for up to at least seven selection rounds, albeit at reduced levels. This enables the discovery of native LNA aptamers and similar oligonucleotide structures. 2012-11-23T09:49:21Z Doessing, Holger og Hanse, Lykke H. og Veedu, Rakesh N. og Wengel, Jesper og Vester, Birte http://espace.library.uq.edu.au/view/UQ:132554 A study of the amplified spontaneous emission (ASE) properties of three bisfluorene-cored dendrimers in the solid state is reported. The results show that the dendron type has a strong impact on the photoluminescence quantum yield and affects the ASE threshold, the optical gain, and loss coefficients. Optically pumped distributed feedback lasers operating in the blue spectral region were fabricated by spin coating the dendrimer films on top of a two-dimensional corrugated fused silica substrate. A best lasing threshold of 4.5 mu J/cm(2) and a slope efficiency of 8.3% were obtained, which demonstrate the high potential of these materials for laser applications. (c) 2007 American Institute of Physics. 2008-03-18T00:00:00Z Ribierre, J. C. og Tsiminis, G. og Richardson, S. og Turnbull, G. A. og Samuel, I. D. W. og Barcena, H. og Burn, P. http://espace.library.uq.edu.au/view/UQ:82170 2007-08-15T00:00:00Z Sly, L. I. http://espace.library.uq.edu.au/view/UQ:104695 2007-08-23T00:00:00Z Ridge, J. P. og Lin, M. og Larsen, E. og Fegan, M. og McEwan, A. G. og Sly, L. I. http://espace.library.uq.edu.au/view/UQ:128170 Pedomicrobium sp. ACM 3067 is a budding-hyphal bacterium belonging to the alpha-Proteobacteria which is able to oxidize soluble Mn2+ to insoluble manganese oxide. A cosmid, from a whole-genome library, containing the putative genes responsible for manganese oxidation was identified and a primer-walking approach yielded 4350 bp of novel sequence. Analysis of this sequence showed the presence of a predicted three-gene operon, moxCBA. The moxA gene product showed homology to multicopper oxidases (MCOs) and contained the characteristic four copper-binding motifs (A, B, C and D) common to MCOs. An insertion mutation of moxA showed that this gene was essential for both manganese oxidation and laccase-like activity. The moxB gene product showed homology to a family of outer membrane proteins which are essential for Type I secretion in Gram-negative bacteria. moxBA has not been observed in other manganese-oxidizing bacteria but homologues were identified in the genomes of several bacteria including Sinorhizobium meliloti 1021 and Agrobacterium tumefaciens C58. These results suggest that moxBA and its homologues constitute a family of genes encoding an MCO and a predicted component of the Type I secretion system. 2008-02-18T00:00:00Z Ridge, Justin P. og Lin, Marianne og Larsen, Eloise I. og Fegan, Mark og McEwan, Alastair G. og Sly, Lindsay I. http://espace.library.uq.edu.au/view/UQ:244337 2011-07-19T00:00:00Z Rothnagel, J. A. og Fisher, M. P. og Axtell, S. M. og Pittelkow, M. R. og Antonlamprecht, I. og Huber, M. og Hohl, D. og Roop, D. R. http://espace.library.uq.edu.au/view/UQ:252660 2011-09-21T00:07:46Z Singh, Yogendra og Sharpe, Philip C. og Hoang, Huy N. og Lucke, Andrew J. og McDowall, Alasdair W. og Bottomley, Stephen P. og Fairlie, David P. http://espace.library.uq.edu.au/view/UQ:262926 Alzheimer’s disease is a leading cause of age-related dementia that is characterized by an extensive loss of neurons and synaptic transmission. The pathological hallmarks of AD are: neurofibrillary tangles and deposition of β-amyloid (Aβ) plaques. Previous research has investigated how Aβ fragments disrupt synaptic mechanisms in the vulnerable regions of the brain. There is a tremendous potential for stem cell technology to extend upon this research not only in terms of developing therapeutic applications, but also in modeling AD. Indeed, the advent of induced pluripotent stem cell technology has opened up exciting new avenues for generating patient and disease-specific cell lines from somatic cells that may be used to model AD. Amyloid precursor protein (APP) is a key protein in neuronal development and this article reviews the role of APP in AD. Stem cell technology offers the opportunity to make use of APP in the directed differentiation of induced pluripotent stem cells into functional neurons, a process that may help generate a model of AD and thereby facilitate an understanding of the mechanisms underlying this disease. 2011-12-05T10:30:17Z Khandekar, Neeta og Lie, Kuhn og Sachdev, Perminder og Sidhu, Kuldip S. http://espace.library.uq.edu.au/view/UQ:255303 2011-10-12T17:51:18Z Counago, Rafael og Wilson, Corey J. og Pena, Matthew I. og Wittung-Stafshede, Pernilla og Shamoo, Yousif http://espace.library.uq.edu.au/view/UQ:64737 Recently, two fresh water species, 'Candidatus Brocadia anammoxidans' and 'Candidatus Kuenenia stuttgartiensis', and one marine species, 'Candidatus Scalindua sorokinii', of planctomycete anammox bacteria have been identified. 'Candidatus Scalindua sorokinii' was discovered in the Black Sea, and contributed substantially to the loss of fixed nitrogen. All three species contain a unique organelle-the anammoxosome-in their cytoplasm. The anammoxosome contains the hydrazine/hydroxylamine oxidoreductase enzyme, and is thus the site of anammox catabolism. The anammoxosome is surrounded by a very dense membrane composed almost exclusively of linearly concatenated cyclobutane-containing lipids. These so-called 'ladderanes' are connected to the glycerol moiety via both ester and ether bonds. In natural and man-made ecosystems, anammox bacteria can cooperate with aerobic ammonium-oxidising bacteria, which protect them from harmful oxygen, and provide the necessary nitrite. The cooperation of these two groups of ammonium-oxidising bacteria is the microbial basis for a sustainable one reactor system, CANON (completely autotrophic nitrogen-removal over nitrite) to remove ammonia from high strength wastewater. 2007-08-14T19:21:51Z Jetten, M. S. og Sliekers, O. og Kuypers, M. og Dalsgaard, T. og van Niftrik, L. og Cirpus, I. og van de Pas-Schoonen, K. T. og Lavik, G. og Thamdrup, B. og Le Paslier, D. og Op den Camp, H. J. M. og Huth, S. og Nielsen, L. P. og Abma, W. og Third, K. og Engstrom, P. og Kuenen, J. G. og Jorgensen, B. B. og Canfield, D. E. og Damste, J. S. S. og Revsbech, N. P. og Fuerst, J. og Weissenbach, J. og Wagner, M. og Schmidt, I. og Schmid, M. og Strous, M. http://espace.library.uq.edu.au/view/UQ:183631 2009-09-04T10:26:51Z Burow, Luke C. og Mabbett, Amanda N. og Borras, Luis og Blackall, Linda L. http://espace.library.uq.edu.au/view/UQ:204671 2010-05-02T00:06:30Z Falsetta, ML og McEwan, AG og Jennings, MP og Apicella, MA http://espace.library.uq.edu.au/view/UQ:144426 2008-06-10T00:00:00Z Bond, PL og Keller, J og Blackall, LL http://espace.library.uq.edu.au/view/UQ:110056 Cytokine mRNA levels were assessed in Burkholderia pseudomallei-suseeptible BALB/c mice and B. pseudomallei-resistant C57BL/6 mice following administration of a sublethal dose of less virulent (LV) B. pseudomallei, a candidate immunogen tested for protection against a highly virulent (HV) challenge. Compared on the basis of the bacterial loads, the cytokine patterns induced by RV and LV B. pseudomallei were similar, involving gamma interferon, interleukin-10, and other cytokines. Partial cross-protection between B. pseudomallei strains is shown to be associated with cytokine profiles involving both type 1 and type 2 cytokines. 2007-09-19T00:00:00Z Ulett, G. C. og Ketheesan, N. og Clair, T. W. og McElnea, C. L. og Barnes, J. L. og Hirst, R. G. http://espace.library.uq.edu.au/view/UQ:62082 2007-08-14T00:00:00Z Winzor, D. J. http://espace.library.uq.edu.au/view/UQ:34764 A simple theoretical framework is presented for bioassay studies using three component in vitro systems. An equilibrium model is used to derive equations useful for predicting changes in biological response after addition of hormone-binding-protein or as a consequence of increased hormone affinity. Sets of possible solutions for receptor occupancy and binding protein occupancy are found for typical values of receptor and binding protein affinity constants. Unique equilibrium solutions are dictated by the initial condition of total hormone concentration. According to the occupancy theory of drug action, increasing the affinity of a hormone for its receptor will result in a proportional increase in biological potency. However, the three component model predicts that the magnitude of increase in biological potency will be a small fraction of the proportional increase in affinity. With typical initial conditions a two-fold increase in hormone affinity for its receptor is predicted to result in only a 33% increase in biological response. Under the same conditions an Ii-fold increase in hormone affinity for receptor would be needed to produce a two-fold increase in biological potency. Some currently used bioassay systems may be unrecognized three component systems and gross errors in biopotency estimates will result if the effect of binding protein is not calculated. An algorithm derived from the three component model is used to predict changes in biological response after addition of binding protein to in vitro systems. The algorithm is tested by application to a published data set from an experimental study in an in vitro system (Lim et al., 1990, Endocrinology 127, 1287-1291). Predicted changes show good agreement (within 8%) with experimental observations. (C) 1998 Academic Press Limited. 2007-08-13T00:00:00Z Barnard, R. T. og Wolff, R. C. http://espace.library.uq.edu.au/view/UQ:98137 2007-08-24T00:00:00Z Zalucki, Y. M. og Terry, T. D. og Blackall, P. J. og Jennings, M. P. http://espace.library.uq.edu.au/view/UQ:111857 The establishment of persistent noncytopathic replication by replicon RNAs of a number of positive-strand RNA viruses usually leads to generation of adaptive mutations in nonstructural genes. Some of these adaptive mutations (e.g., in hepatitis C virus) increase the ability of RNA replication to resist the antiviral action of alpha/beta interferon (IFN-alpha/beta); others (e.g., in Sindbis virus) may also lead to more efficient IFN production. Using puromycin-selectable Kunjin virus (KUN) replicon RNA, we identified two adaptive mutations in the NS2A gene (producing Ala30-to-Pro and Asn101-to-Asp mutations in the gene product; for simplicity, these will be referred to hereafter as Ala30-to-Pro and Asn101-to-Asp mutations) that, when introduced individually or together into the original wild-type (wt) replicon RNA, resulted in similar to15- to 50-fold more efficient establishment of persistent replication in hamster (BHK21) and human (HEK293 and HEp-2) cell lines. Transfection with a reporter plasmid carrying the luciferase gene under the control of the IFN-beta promoter resulted in similar to6- to 7-fold-higher luciferase expression in HEp-2 cells stably expressing KUN replicon RNA with an Ala30-to-Pro mutation in the NS2A gene compared to that observed in HEp-2 cells stably expressing KUN replicon RNA with the wt NS2A gene. Moreover, cotransfection of plasmids expressing individual wt or Ala30-to-Pro-mutated NS2A genes with the IFN-beta promoter reporter plasmid, followed by infection with Semliki Forest virus to activate IFN-beta promoter-driven transcription, showed similar to7-fold inhibition of luciferase expression by the wt but not by the Ala30-to-Pro-mutated NS2A protein. The results show for the first time a role for the flavivirus nonstructural protein NS2A in inhibition of IFN-beta promoter-driven transcription and identify a single-amino-acid mutation in NS2A that dramatically reduces this inhibitory activity. The findings determine a new function for NS2A in virus-host interactions, extend the range of KUN replicon vectors for noncytopathic gene. expression, and identify NS2A as a new target for attenuation in the development of live flavivirus vaccines. 2007-09-19T00:00:00Z Liu, Wen Jun og Chen, Hua Bo og Xiang, Ju Wang og Huang, Hester og Khromykh, Alexander A. http://espace.library.uq.edu.au/view/UQ:98142 2007-08-24T00:00:00Z Hobb, Rhonda I. og Tseng, Hsing-Ju og Downes, John E. og Terry, Tamsin D. og Blackall, Patrick J. og Jennings, Michael P. http://espace.library.uq.edu.au/view/UQ:283010 2012-10-08T15:50:33Z Chan, Cheong Xin og Soares, Marcelo B. og Bonaldo, Maria F. og Wisecaver, Jennifer H. og Hackett, Jeremiah D. og Anderson, Donald M. og Erdner, Deana L. og Bhattacharya, Debashish http://espace.library.uq.edu.au/view/UQ:272996 2012-04-23T08:30:36Z Ben Zakour, Nouri L. og Venturini, Carola og Beatson, Scott A. og Walker, Mark J. http://espace.library.uq.edu.au/view/UQ:229791 2011-02-22T00:00:00Z Smart, C.E. og Wronski, A. og French, J.D. og Edwards, S.L. og Asselin-Labat, M.L. og Waddell, N. og Peters, K. og Brewster, B.L. og Brooks, K. og Simpson, K. og Manning, N. og Lakhani, S.R. og Grimmond, S. og Lindeman, G.J. og Visvader, J.E. og Brown, M.A. http://espace.library.uq.edu.au/view/UQ:283615 2012-10-19T10:22:17Z Bailey, Ulla-Maja og Jamaluddin, Muhammad Fairuz og Schulz, Benjamin L. http://espace.library.uq.edu.au/view/UQ:231668 2011-03-07T00:00:00Z Chow, Sharon og Fletcher, Mary T. og Mckenzie, Ross A. http://espace.library.uq.edu.au/view/UQ:267843 2012-02-18T00:00:00Z Chow, Sharon og Fletcher, Mary T. og Mckenzie, Ross A. http://espace.library.uq.edu.au/view/UQ:62410 Genetic diversity in Cassia brewsteri (F. Muell.) F. Muell. ex Benth. was assessed with Randomly Amplified DNA Fingerprints (RAFs). Thirty accessions of C. brewsteri collected from throughout its natural distribution were analysed with three random decamer primers, along with three accessions of C. tomentella (Benth.) Domin and a single accession of each of C. queenslandica C. T. White and C. marksiana (F. M. Bailey) Domin. The three primers yielded a reproducible amplification profile of 265 scorable polymorphic fragments for the 35 accessions. These molecular markers were used to calculate Nei and Li similarity coefficients between each pair of individuals. A matrix of dissimilarity of each pair of individuals was examined by multidimensional scaling (MDS). The analysis supports the division of C. brewsteri into two subspecies and the suggestion that intergradation of C. brewsteri and C. tomentella can occur where the distributions of these species meet. 2007-08-14T00:00:00Z Cunningham, D. C. og Walsh, K. B. og Anderson, E. R. og Harrison, D. K. og Carroll, B. J. http://espace.library.uq.edu.au/view/UQ:217512 Asparagine-linked glycosylation is the most common post-translational modification of proteins catalyzed in eukaryotes by the multiprotein complex oligosaccharyltransferase. Apart from the catalytic Stt3p, the roles of the subunits are ill defined. Here we describe functional investigations of the Ost3/6p components of the yeast enzyme. We developed novel analytical tools to quantify glycosylation site occupancy by enriching glycoproteins bound to the yeast polysaccharide cell wall, tagging glycosylated asparagines using endoglycosidase H glycan release, and detecting peptides and glycopeptides with LC-ESI-MS/MS. We found that the paralogues Ost3p and Ost6p were required for efficient glycosylation of distinct defined glycosylation sites. Our results describe a novel method for relative quantification of glycosylation occupancy in the genetically tractable yeast system and show that eukaryotic oligosaccharyltransferase isoforms have different activities toward protein substrates at the level of individual glycosylation sites. 2010-09-30T00:00:00Z Schulz, Benjamin L. og Aebi, Markus http://espace.library.uq.edu.au/view/UQ:36855 The analysis of keratin 6 expression is complicated by the presence of multiple isoforms that are expressed constitutively in a number of internal stratified epithelia, in palmoplantar epidermis, and in the companion cell layer of the hair follicle. In addition, keratin 6 expression is inducible in interfollicular epidermis and the outer root sheath of the follicle, in response to wounding stimuli, phorbol esters, or retinoic acid. In order to establish the critical regions involved in the regulation of keratin 6a (the dominant isoform in mice), we generated transgenic mice with two different-sized mouse keratin 6a constructs containing either 1.3 kb or 0.12 kb of 5' flanking sequence linked to the lacZ reporter gene. Both constructs also contained the first intron and the 3' flanking sequence of mouse keratin 6a. Ectopic expression of either transgene was not observed. Double-label immunofluorescence analyses demonstrated expression of the reporter gene in keratin 6 expressing tissues, including the hair follicle, tongue, footpad, and nail bed, showing that both transgenes retained keratinocyte-specific expression. Quantitative analysis of beta -galactosidase activity verified that both the 1.3 and 0.12 kb keratin 6a promoter constructs produced similar levels of the reporter. Notably, both constructs were constitutively expressed in the outer root sheath and interfollicular epidermis in the absence of any activating stimulus, suggesting that they lack the regulatory elements that normally silence transcription in these cells. This study has revealed that a keratin 6a minigene contains critical cis elements that mediate tissue-specific expression and that the elements regulating keratin 6 induction lie distal to the 1.3 kb promoter region. 2007-08-13T00:00:00Z Mahony, D. og Karunaratne, S. M. og Cam, G. og Rothnagel, J. A. http://espace.library.uq.edu.au/view/UQ:267294 2012-02-09T10:15:35Z Issa, Samah M. A. og Schulz, Benjamin L. og Packer, Nicolle H. og Karlsson, Niclas G. http://espace.library.uq.edu.au/view/UQ:128238 Fluorescence-based PCR techniques are becoming an increasingly popular method for measuring low-abundance alternatively spliced mRNA transcripts. The dynamic range of real-time RT-PCR affords high sensitivity for the measurement of gene expression, but this mandates the need for strict controls to ensure assay validity. Primer design, reverse transcription, and cycling conditions need to be optimized to ensure an accurate and reproducible assay. Here, we describe a procedure for creating a cost effective and reliable method for the absolute quantification of several exon-skipping variants of human excitatory amino acid transporter-2 (EAAT2). We show that the cycling conditions can be adjusted to increase the specificity of primers that span exon-exon junctions, and that differences in the reverse transcription reaction can be minimized. Standard curves are stable and produce accurate absolute copy number data. We report that exon-skipping transcripts, EAAT2 Delta 7 and EAAT2 Delta 9, account for 5.8% of EAAT2 mRNA in autopsy human neocortex. (c) 2006 Elsevier B.V. All rights reserved. 2008-02-18T00:00:00Z Walton, Heather S. og Gebhardt, Florian M. og Innes, David J. og Dodd, Peter R. http://espace.library.uq.edu.au/view/UQ:219005 2010-10-24T00:10:45Z Munce, T. og Heussler, H. S. og Bowling, F. G. http://espace.library.uq.edu.au/view/UQ:150594 2008-06-06T00:00:00Z Heelan, L. A. og Bassam, B. J. og Croft, B. J. og Dietzgen, R. G. og Maclean, D. J. http://espace.library.uq.edu.au/view/UQ:273810 2012-05-10T15:18:08Z Chan, Cheong Xin og Zauner, Simone og Wheeler, Glen og Grossman, Arthur R. og Prochnik, Simon E. og Blouin, Nicolas A. og Zhuang, Yunyun og Benning, Christoph og Berg, Gry Mine og Yarish, Charles og Eriksen, Renee L. og Klein, Anita S. og Lin, Senjie og Levine, Ira og Brawley, Susan H. og Bhattacharya, Debashish http://espace.library.uq.edu.au/view/UQ:243312 We are developing a system for isolating tomato genes by transposon mutagenesis. In maize and tobacco, the transposon Activator (Ac) transposes preferentially to genetically linked sites. To identify transposons linked to various target genes, we have determined the RFLP map locations of Ac- and Dissociation (Ds)-carrying T-DNAs in a number of transformants. T-DNA flanking sequences were isolated using the inverse polymerase chain reaction (IPCR) and located on the RFLP map of tomato. The authenticity of IPCR reaction products was tested by several criteria including nested primer amplification, DNA sequence analysis and PCR amplification of the corresponding insertion target sequences. We report the RFLP map locations of 37 transposon-carrying T-DNAs. We also report the map locations of nine transposed Ds elements. T-DNAs were identified on all chromosomes except chromosome 6. Our data revealed no apparent chromosomal preference for T-DNA integration events. Lines carrying transposons at known map locations have been established which should prove a useful resource for isolating tomato genes by transposon mutagenesis. 2011-07-07T09:29:30Z Thomas, Colwyn M. og Jones, David A. og English, James J. og Carroll, Bernard J. og Bennetzen, Jeffrey L. og Harrison, Kate og Burbidge, Alan og Bishop, Gerard J. og Jones, Jonathan D. G. http://espace.library.uq.edu.au/view/UQ:290011 Saliva is a crucial biofluid for oral health and is also of increasing importance as a non-invasive source of disease biomarkers. Salivary alpha-amylase is an abundant protein in saliva, and changes in amylase expression have been previously associated with a variety of diseases and conditions. Salivary alpha-amylase is subject to a high diversity of post-translational modifications, including physiological proteolysis in the oral cavity. Here we developed methodology for rapid sample preparation and non-targeted LC-ESI-MS/MS analysis of saliva from healthy subjects and observed an extreme diversity of alpha-amylase proteolytic isoforms. Our results emphasize the importance of consideration of post-translational events such as proteolysis in proteomic studies, biomarker discovery and validation, particularly in saliva. 2013-01-27T00:10:09Z Bailey, Ulla-Maja og Punyadeera, Chamindie og Cooper-White, Justin J. og Schulz, Benjamin L. http://espace.library.uq.edu.au/view/UQ:218306 2010-10-12T00:00:00Z Byrne L.T. og Ferro, V. og Stevenson, S. og Stick, R.V. http://espace.library.uq.edu.au/view/UQ:176890 2009-04-17T16:28:04Z Aaron Poth og Deeth, Hilton C. og Alewood, Paul F. og Holland, John W.