Open Access. Powered by Scholars. Published by Universities.®

Life Sciences Commons

Open Access. Powered by Scholars. Published by Universities.®

Articles 1 - 11 of 11

Full-Text Articles in Life Sciences

Expression And Function Of Alpha3 And Beta2 Neuronal Nicotinic Acetylcholine Receptor Subunits In Hek-293 Cells, Nathan W. Steinhafel Dec 2006

Expression And Function Of Alpha3 And Beta2 Neuronal Nicotinic Acetylcholine Receptor Subunits In Hek-293 Cells, Nathan W. Steinhafel

Theses and Dissertations

Single-cell real-time quantitative RT-PCR was used to characterize the mRNA expression of rat neuronal nicotinic acetylcholine receptor (nAChR) subunits α3 and β2 in CA1 hippocampus stratum radiatum and stratum oriens interneurons. α3β2 co-expression was detected in 43% of interneurons analyzed. The nAChR subtype α3β2 was transiently expressed in cells derived from the human embryonic kidney cell line 293 at mRNA levels found in the CA1. The functional properties of α3β2 in HEK-293 cells were characterized by whole-cell patch clamping using acetylcholine (ACh) as an agonist. The kinetics of α3β2 channels were further analyzed by altering the level of α3 DNA …


Biophotonics: Electrochemiluminescence At Microelectrodes During Pcr Amplification Of Dna, Rosemary L. Smith, Scott Collins Aug 2006

Biophotonics: Electrochemiluminescence At Microelectrodes During Pcr Amplification Of Dna, Rosemary L. Smith, Scott Collins

University of Maine Office of Research Administration: Grant Reports

This project investigates a new technique for in situ quantification of Polymerase Chain Reaction (PCR) amplification products using electrochemiluminescence (ECL). The technique implements the highly sensitive, ECL detection and quantification of tris(2,2'bipyridyl) ruthenium (II) labeled DNA. This method is expected to yield significant improvement in speed, cost and performance over existing quantitative PCR methods, by reducing the number and quantity of reagents, reducing the number of sample preparation steps, increasing sensitivity, and shortening analysis time.


The Lift Pool Method For Isolation Of Cdna Clones From Lambda Phage Libraries, Janine M. Leblanc-Straceski, Pablo Sobrado, Sharon Betz, Julie Zerfas, Karen Morgan Jul 2006

The Lift Pool Method For Isolation Of Cdna Clones From Lambda Phage Libraries, Janine M. Leblanc-Straceski, Pablo Sobrado, Sharon Betz, Julie Zerfas, Karen Morgan

Biology Faculty Publications

A PCR based strategy was developed to screen a Xenopus oocyte λgt10 cDNA library. The PCR-based lift pool (LP) method follows the same two tiered strategy as conventional screening of phage libraries by filter hybridization. Two rounds of plating, one at high density to detect the clone, and one at low density to purify the clone to homogeneity, are performed. In the first round, lysates from high density plates, termed plate pools (PP), serve as template for PCR. In the second round, phage particles adhering to plaque lifts of low density plates are washed off nitrocellulose filters to create LPs, …


Bacillus Anthracis Virulent Plasmid Px02 Genes Found In Large Plasmids Of Two Other Bacillus Species, Vicki A. Luna, Debra S. King, K. Kealy Peak, Frank Reeves, Lea Heberlein-Larson, William Veguilla, L. Heller, Kathleen E. Duncan, Andrew C. Cannons, Philip Amuso, Jacqueline Cattani Jul 2006

Bacillus Anthracis Virulent Plasmid Px02 Genes Found In Large Plasmids Of Two Other Bacillus Species, Vicki A. Luna, Debra S. King, K. Kealy Peak, Frank Reeves, Lea Heberlein-Larson, William Veguilla, L. Heller, Kathleen E. Duncan, Andrew C. Cannons, Philip Amuso, Jacqueline Cattani

Bioelectrics Publications

In order to cause the disease anthrax, Bacillus anthracis requires two plasmids, pX01 and pX02, which carry toxin and capsule genes, respectively, that are used as genetic targets in the laboratory detection of the bacterium. Clinical, forensic, and environmental samples that test positive by PCR protocols established by the Centers for Disease Control and Prevention for B. anthracis are considered to be potentially B. anthracis until confirmed by culture and a secondary battery of tests. We report the presence of 10 genes (acpA, capA, capB, capC, capR, capD, IS1627, ORF 48, ORF 61, and repA) and the sequence for the …


Comparison Of Polymerase Chain Reaction And Conventional Methods For The Diagnosis Of Listeria Monocytogenes In Stuffed Mussels, Ergün Ö. Göksoy, Şükrü Kirkan, Osman Kaya Jan 2006

Comparison Of Polymerase Chain Reaction And Conventional Methods For The Diagnosis Of Listeria Monocytogenes In Stuffed Mussels, Ergün Ö. Göksoy, Şükrü Kirkan, Osman Kaya

Turkish Journal of Veterinary & Animal Sciences

The aim of this study was to evaluate the role of stuffed mussel as a source of Listeria monocytogenes. Polymerase chain reaction is a rapid procedure with both sensitivity and specificity for quick detection and identification of L. monocytogenes. A total of 50 mussel samples were investigated for L. monocytogenes. L. monocytogenes was not identified by the conventional method. However, PCR amplification products demonstrated that 5 out of 50 samples showed positive reactions with L. monocytogenes. The PCR positive samples showed specific amplification at approximately 343 bp for L. monocytogenes.


Isolation And Characterization Of Toxin A-Negative, Toxin B-Positive Clostridium Difficile In Dublin, Ireland, Denise Drudy, N. Harnedy, S. Fanning, R. O’Mahony, L. Kyne Jan 2006

Isolation And Characterization Of Toxin A-Negative, Toxin B-Positive Clostridium Difficile In Dublin, Ireland, Denise Drudy, N. Harnedy, S. Fanning, R. O’Mahony, L. Kyne

Articles

Clostridium difficile is a major cause of infectious diarrhoea in hospitalised patients. Most pathogenic C. difficile strains produce two toxins, A and B; however, clinically relevant toxin A-negative, toxin Bpositive (A– B+ ) strains of C. difficile that cause diarrhoea and colitis in humans have been isolated worldwide. The aims of this study were to isolate and characterise A– B+ strains from two university hospitals in Dublin, Ireland. Samples positive for C. difficile were identified daily by review of ELISA results and were cultured on selective media. Following culture, toxin-specific immunoassays, IMR-90 cytotoxicity assays and PCR were used to analyse …


Identification Of Vibrio Anguillarum By Pcr (Rpon Gene) Associated With Vibriosis In Marine Fish In Turkey, Di̇dem Demi̇rcan, Akin Candan Jan 2006

Identification Of Vibrio Anguillarum By Pcr (Rpon Gene) Associated With Vibriosis In Marine Fish In Turkey, Di̇dem Demi̇rcan, Akin Candan

Turkish Journal of Veterinary & Animal Sciences

The main causative agent of vibriosis is Vibrio anguillarum. In this study, 33 V. anguillarum strains were isolated from the internal organs of marine fish showing typical clinical signs of vibriosis, and these strains were identified by biochemical tests. For comparison of these results, API 20E and BIONOR Mono-Va agglutination kits were used. Finally strains were amplified by PCR using the rpoN gene. Although the phenotypic properties of V. anguillarum strains varied, the rpoN gene from chromosomal DNA was amplified in all strains. In conclusion, this gene could be used in the diagnosis of V. anguillarum strains isolated in Turkey.


Quantification Of Mrna Expression Of Her2/Neu, Estrogen Receptors Eralpha And Erbeta, And Progesterone Receptor Using Real-Time Reverse Transcription Pcr In Small Tumor Biopsies, In Human Breast Cancer, Evonne Marie. Mc Cabe Jan 2006

Quantification Of Mrna Expression Of Her2/Neu, Estrogen Receptors Eralpha And Erbeta, And Progesterone Receptor Using Real-Time Reverse Transcription Pcr In Small Tumor Biopsies, In Human Breast Cancer, Evonne Marie. Mc Cabe

Theses

Early detection is an effective means of reducing cancer mortality. Early detection greatly improves treatment options and the chances for successful treatment. The prognosis for patients with breast cancer is determined by well-established biological features associated with biological aggressiveness, histological grade, tumour size, and nodal involvement. Breast cancer results from multiple genetic abnormalities, and cellular differentiation, tumour growth, and metastatic potential are the end result of these changes. Identification of molecular changes is of considerable value in enhancing the accuracy of prognostic indices, and more importantly, it may indicate various changes as putative targets for therapy. In recent years a …


Detection Of Listeria Monocytogenes By Using Pcr In Helix Pomatia, Şükrü Kirkan, Ergün Ö. Göksoy, Osman Kaya Jan 2006

Detection Of Listeria Monocytogenes By Using Pcr In Helix Pomatia, Şükrü Kirkan, Ergün Ö. Göksoy, Osman Kaya

Turkish Journal of Veterinary & Animal Sciences

The detection of Listeria monocytogenes in Helix pomatia, both raw and cooked is presented here. Polymerase chain reaction (PCR) is a rapid procedure with both sensitivity and specificity for quick detection and identification of Listeria monocytogenes from raw and cooked Helix pomatia. A total of 30 bags (10 g each) of H. pomatia samples were investigated for the Listeria monocytogenes inlB gene with the PCR method. PCR amplification products demonstrated that 18 of the 30 samples showed positive reactions to Listeria monocytogenes in the PCR. All PCR positive samples showed specific amplification of the 343 bp fragment for Listeria monocytogenes.


Characterization Of Footrot Bacteria Dichelobacter Nodosus Using Pcr Amplification And Dna Sequence Analysis, Ifakat Tülay Çağatay, Jon G. H. Hickford Jan 2006

Characterization Of Footrot Bacteria Dichelobacter Nodosus Using Pcr Amplification And Dna Sequence Analysis, Ifakat Tülay Çağatay, Jon G. H. Hickford

Turkish Journal of Veterinary & Animal Sciences

Dichelobacter nodosus is an essential causative agent of footrot in ruminants, particularly in sheep, goats and cattle. In this study, more than 100 footrot samples were collected from 4 different farming regions in New Zealand (NZ). Selective media were chosen and isolation and routine growth conditions were optimized for NZ D. nodosus serotypes. Approximately 1000 primary plates were anaerobically subcultured several times and examined with Gram staining in order to detect single colonies of D. nodosus. Both the variable region and a part of the conserved region of fimbrial subunit gene (fimA) were amplified from bacterial DNA using polymerase chain …


Nucleic Acid Extraction From Clinical Specimens For Pcr Applications, Ashabil Aygan Jan 2006

Nucleic Acid Extraction From Clinical Specimens For Pcr Applications, Ashabil Aygan

Turkish Journal of Biology

Polymerase chain reaction (PCR) has been widely used due to its high specificity, sensitivity, and rapid turn-around time. However, the major limitation of PCR based diagnosis is inhibition of the reaction caused by a variety of components within clinical specimens. The demand for PCR diagnosis in medical microbiology has highlighted the need for efficient methods of nucleic acid extraction. Although there has been progress in the simplification and purification of microbial nucleic acids, many researchers' procedures are still inconvenient for routine use with any specimen and a universal method has not been devised yet. In this review, some of the …