• Title/Summary/Keyword: novel primer

Search Result 100, Processing Time 0.028 seconds

Fabrication and Characterization of Novel Electrospun PVPA/PVA Nanofiber Matrix for Bone Tissue Engineering

  • Franco, Rose-Ann;Nguyen, Thi Hiep;Lee, Byong-Taek
    • Proceedings of the Materials Research Society of Korea Conference
    • /
    • 2011.05a
    • /
    • pp.51.2-51.2
    • /
    • 2011
  • A novel electrospun nanofiber membrane was fabricated using combined poly (vinylphosphonic acid) (PVPA) and polyvinyl alcohol (PVA) intended for bone tissue engineering applications. PVPA is a proton-conducting polymer used as primer for bone implants and dental cements to prevent corrosion and brush abrasion. The phosphonate groups of PVPA have the ability to crosslink and attach itself to the hydroxyapatite surface facilitating faster integration of the biomaterial to the bone matrix. PVA was combined with PVPA to provide hydrophilicity, biocompatibility and improve its spinnability. To improve its mechanical strength, PVPA/PVA and neat PVA mixtures were combined to produce a multilayer scaffold. The physical and chemical properties of the of the fabricated matrix was investigated by SEM and TEM morphological analyses, tensile strength test, XRD, FT-IR spectra, swelling behavior and biodegradation rates, porosity and contact angle measurements. Biocompatibility was also examined in vitro by cytotoxicity and cell proliferation studies with MTT assay and cell adhesion behavior by SEM and confocal microscopy.

  • PDF

Mycoplasma hyopneumoniae Induces Grap, Gadd45β, and secreted phosphoprotein 1 Gene Expression as Part of the Inflammatory Response in RAW264.7 Cells

  • Hwang, Mi-Hyun;Choi, Myung-Jin;Park, Seung-Chun
    • Toxicological Research
    • /
    • v.25 no.3
    • /
    • pp.119-124
    • /
    • 2009
  • Genes related to Mycoplasma hyopneumoniae-induced inflammation were identified using the genefishing technology, an improved method for identifying differentially expressed genes (DEGs) using an annealing control primer (ACP) system in RAW264.7 cells. After treatment with M. hyopneumoniae, 16 DEGs were expressed in RAW264.7 cells using a pre-screening system. Among these 16 DEGs, 11 DEGs (DEGs 1, 4, 5-10, 12-15) were selected and sequenced directly, revealing that DEG12 (Grap), DEG14 (Gadd45), and DEG15 (secreted phosphoprotein 1) were related to inflammatory cytokines. This is the first report that intact M. hyopneumoniae induces the expression of Grap, Gadd 45${\beta}$, and secreted phosphoprotein 1 in RAW264.7 cells. Subsequently, these genes may be targets for screening novel inhibitors of the mycoplasmal inflammatory response.

Preparation and Characterization of Coating Solution Based on Waterborne Polyurethane Dispersion containing Fluorine for Primer on Electro Galvanized Steel Sheet

  • Jin, Chung Keun;Lim, Sung Hyung
    • Corrosion Science and Technology
    • /
    • v.14 no.5
    • /
    • pp.207-212
    • /
    • 2015
  • The purpose of this research was to synthesize fluorine modified waterborne polyurethane dispersion (F-WPU) by soap-free (internal emulsifier) emulsion polymerization techniques, to prepare coating solution based on fluorine modified waterborne polyurethane dispersion (F-WPU) and to compare the chemical and thermo-mechanical properties on the electrogalvanized steel sheet. Environmentally friendly F-WPU was prepared with a fluorinated polyol containing 60 wt% of fluorine. There are various ways of combining a wide variety of fluorinated polyols and diisocyanate to exhibit novel properties of waterborne polyurethane dispersion. Components of coating solution were largely divided into 4 kinds i.e., F-WPU, acrylic emulsion, silane coupling agent, and colloidal silicate. F-WPU coating solution on the electro-galvanized steel sheet showed excellent properties of corrosion resistance, alkali resistance and heat resistance, as compared to other coating solutions using a general waterborne resin. The F-WPU coating solution's reliable effects are possibly due to the fluorine atoms incorporated even in a small amount of F-WPU.

Corrosion Protection of Automotive Steels by Novel Water-borne Primer Systems

  • Ooij, William J. van;Puomi, Paula
    • Corrosion Science and Technology
    • /
    • v.6 no.5
    • /
    • pp.239-244
    • /
    • 2007
  • Corrosion protection of automotive steels has traditionally been assured by using a zinc phosphate metal pretreatment followed by the deposition of a cathodic electrocoat system. This system has been developed and optimized over the years into a highly robust and dependable system with a high performance. However, in terms of efficiency and use of resources and energy, the need is now felt to develop a simpler system with fewer steps, shorter lines, less energy requirements (curing and e-coat deposition) and less stringent waste disposal requirement (phosphate sludge). We report here on the development of a one-step system that can possibly replace both the zinc phosphate and the e-coating processes. Such a system is based on the so-called superprimer concept that we have recently developed for the replacement of chromate pretreatment and chromate-containing primers in the aerospace industry. With some modifications, such systems can also be adapted for use in the automotive industry.

Characterization of the xaiF Gene Encoding a Novel Xylanase-activity- increasing Factor, XaiF

  • Cho, Ssang-Goo;Choi, Yong-Jin
    • Journal of Microbiology and Biotechnology
    • /
    • v.8 no.4
    • /
    • pp.378-387
    • /
    • 1998
  • The DNA sequence immediately following the xynA gene of Bacillus stearothermophilus 236 [about l-kb region downstream from the translational termination codon (TAA) of the xynA gene]was found to have an ability to enhance the xylanase activity of the upstream xynA gene. An 849-bp ORF was identified in the downstream region, and the ORF was confirmed to encode a novel protein of 283 amino acids designated as XaiF (xylanase-activity-increasing factor). From the nucleotide sequence of the xaiF gene, the molecular mass and pI of XaiF were deduced to be 32,006 Da and 4.46, respectively. XaiF was overproduced in the E. coli cells from the cloned xaiF gene by using the T7 expression system. The transcriptional initiation site was determined by primer extension analysis and the putative promoter and ribosome binding regions were also identified. Blast search showed that the xaiF and its protein product had no homology with any gene nor any protein reported so far. Also, in B. subtilis, the xaiF trans-activated the xylanase activity at the same rate as in E. coli. In contrast, xaiF had no activating effect on the co-expressed ${\beta}-xylosidase$ of the xylA gene derived from the same strain of B. stearothermophilus. In addition, the intracellular and extracellular fractions from the E. coli cells carrying the plasmid-borne xaiF gene did not increase the isolated xylanase activity, indicating that the protein-protein interaction between XynA and XaiF was not a causative event for the xylanase activating effect of the xaiF gene.

  • PDF

Rapid Identification of Ginseng Cultivars (Panax ginseng Meyer) Using Novel SNP-Based Probes

  • Jo, Ick-Hyun;Bang, Kyong-Hwan;Kim, Young-Chang;Lee, Jei-Wan;Seo, A-Yeon;Seong, Bong-Jae;Kim, Hyun-Ho;Kim, Dong-Hwi;Cha, Seon-Woo;Cho, Yong-Gu;Kim, Hong-Sig
    • Journal of Ginseng Research
    • /
    • v.35 no.4
    • /
    • pp.504-513
    • /
    • 2011
  • In order to develop a novel system for the discrimination of five ginseng cultivars (Panax ginseng Meyer), single nucleotide polymorphism (SNP) genotyping assays with real-time polymerase chain reaction were conducted. Nucleotide substitution in gDNA library clones of P. ginseng cv. Yunpoong was targeted for the SNP genotyping assay. From these SNP sites, a set of modified SNP specific fluorescence probes (PGP74, PGP110, and PGP130) and novel primer sets have been developed to distinguish among five ginseng cultivars. The combination of the SNP type of the five cultivars, Chungpoong, Yunpoong, Gopoong, Kumpoong, and Sunpoong, was identified as 'ATA', 'GCC', 'GTA', 'GCA', and 'ACC', respectively. This study represents the first report of the identification of ginseng cultivars by fluorescence probes. An SNP genotyping assay using fluorescence probes could prove useful for the identification of ginseng cultivars and ginseng seed management systems and guarantee the purity of ginseng seed.

Cloning and Characterization of Novel Cytochrome P450 Hydroxylase Genes from Pseudonocardia autotrophica (Pseudonocardia autotrophica 유래의 신규 Cytochrome Cytochrome P450 Hydroxylase 유전자의 분리 및 염기서열 특성규명)

  • Myeong Ji Seon;Park Hyun-Joo;Han Kyuboem;Kim Sang-Nyun;Kim Eung-Soo
    • Korean Journal of Microbiology
    • /
    • v.40 no.3
    • /
    • pp.221-225
    • /
    • 2004
  • Novel cytochrome P450 hydroxylase (CYP) genes were isolated and characterized from P. autotrophica cosmid DNA library using an actinomycete CYP-specific PCR product as a screening probe. The cosmids containing four unique CYP genes (pESK601, 602, 603, 604, 605) were identified, and the four CYP genes were completely sequenced to be homologous to other known Actinomycetes CYP genes involved in various secondary metabolic pathways. Among all novel actinomycete CYP genes found in P. autotrophica, the CYP genes present in pESK601 were revealed to be highly homologous to the CYP genes involved in polyene-type amphotericin and nystatin antibiotic biosynthesis. The nucleotide sequences of the CYP flanking region in pESK601 also revealed the polyene-type biosynthetic genes, implying the presence of a cryptic polyene-type antifungal biosynthetic gene cluster in P. autotrophica.

Genetic Variability Based on Randomly Amplified Polymorphic DNA in Kacip Fatimah (Labisia pumila Benth & Hook f) collected from Melaka and Negeri Sembilan States of Malaysia

  • Bhore, Subhash J.;Nurul, A.H.;Shah, Farida H.
    • Journal of Forest and Environmental Science
    • /
    • v.25 no.2
    • /
    • pp.93-100
    • /
    • 2009
  • In Malaysia, Labisia pumila Benth & Hook f, popularly known as 'Kacip Fatimah' has been used traditionally to treat various elements of the woman's health in Malay community. The objective of this study was to develop randomly amplified polymorphic DNA (RAPD) based DNA markers for the identification of L. pumila and to distinguish its three varieties from each other. Total DNA from nine accessions of L. pumila was extracted by CTAB method and polymerase chain reactions (PCR) were carried out to amplify the segments of DNA using different primers to develop DNA barcode using RAPD technique. To find out variety-specific DNA marker/s, twenty different 10-mer primer sequences with annealing temperature from 36-$40^{\circ}C$ were evaluated in triplicate. Out of 20 random primers, two primers (OPA-1 and OPA-2/A10) were selected which produced reliable RAPD band patterns. To have DNA based handle, two RAPD amplification products were cloned and sequenced to determine the identity of the DNA. RAPD analysis using two random primers generated 72 discrete bands ranging in size 200 bp-3,000 bp. Fifty nine of these were polymorphic loci (82%) and thirteen were non-polymorphic loci (18%). A total of 32 bands polymorphic loci (72%) were amplified with primer OPA-1 and analyzed by cluster analysis and UPGMA (Unweighted Pair Group Method with Arithmetic) to present a dendogram depicting the degree of genetic relationship among nine accessions of L. pumila. Our results shows the reasonable genetic diversity among the L. pumila varieties and within varieties; and two RAPD marker sequences obtained could be used to identify L. pumila at species level.

  • PDF

A Simple, Single Triplex PCR of IS6110, IS1081, and 23S Ribosomal DNA Targets, Developed for Rapid Detection and Discrimination of Mycobacterium from Clinical Samples

  • Nghiem, Minh Ngoc;Nguyen, Bac Van;Nguyen, Son Thai;Vo, Thuy Thi Bich;Nong, Hai Van
    • Journal of Microbiology and Biotechnology
    • /
    • v.25 no.5
    • /
    • pp.745-752
    • /
    • 2015
  • Tuberculosis (TB) is the most common mycobacterial infection in developing countries, requiring a rapid, accurate, and well-differentiated detection/diagnosis. For the rapid detection and discrimination of Mycobacterium tuberculosis complex (MTC) from non-tuberculous mycobacteria (NTM), a novel, simple, and primer-combined single-step multiplex PCR using three primer pairs (6110F-6110R, 1081F-1081R, and 23SF-23SR; annealing on each of IS6110, IS1081, and 23S rDNA targets), hereafter referred to as a triplex PCR, has been developed and evaluated. The expected product for IS6110 is 416 bp, for IS1081 is 300 bp, and for 23S rDNA is 206 bp by single PCR, which was used to verify the specificity of primers and the identity of MTC using DNA extracted from the M. tuberculosis H37Rv reference strain (ATCC, USA) and other mycobacteria other than tuberculosis (MOTT) templates. The triplex PCR assay showed 100% specificity and 96% sensitivity; the limit of detection for mycobacteria was ~100 fg; and it failed to amplify any target from DNA of MOTT (50 samples tested). Of 307 blinded clinical samples, overall 205 positive M. tuberculosis samples were detected by single PCR, 142 by conventional culture, and 90 by AFB smear methods. Remarkably, the triplex PCR could subsequently detect 55 positive M. tuberculosis from 165 culture-negative and 115 from 217 AFB smear-negative samples. The triplex PCR, targeting three regions in the M. tuberculosis genome, has proved to be an efficient tool for increasing positive detection/discrimination of this bacterium from clinical samples.

Development of PCR-Based Molecular Marker for Detection of Xanthomonas campestris pv. campestris Race 6, the Causative Agent of Black Rot of Brassicas

  • Afrin, Khandker Shazia;Rahim, Md Abdur;Rubel, Mehede Hassan;Park, Jong-In;Jung, Hee-Jeong;Kim, Hoy-Taek;Nou, Ill-Sup
    • The Plant Pathology Journal
    • /
    • v.36 no.5
    • /
    • pp.418-427
    • /
    • 2020
  • Xanthomonas campestris pv. campestris (Xcc), the pathogen of black rot which is the most destructive disease of Brassica vegetables throughout the world. Here, we reported two novel sequence-characterized amplified region (SCAR) markers (i.e., XccR6-60 and XccR6-67) for the detection of Xcc race 6 via re-alignment of the complete genome sequences of Xcc races/strains/pathovars. The specificity of SCAR primer sets was verified by mean of PCR amplification using the genomic DNA template of Xcc races/strains/pathovars and two other plant infecting bacterial strains. The PCR result revealed that the XccR6-60 and XccR6-67 primer sets amplified 692-bp and 917-bp DNA fragments, respectively, specifically from race 6, while no visible amplification was detected in other samples. In addition, the SCAR primers were highly sensitive and can detect from a very low concentration of genomic DNA of Xcc race 6. However, the complete genome sequence of Xcc race 6 is not yet publicly available. Therefore, the cloning and sequencing of XccR6-60 and XccR6-67 fragments from race 6 provide more evidence of the specificity of these markers. These results indicated that the newly developed SCAR markers can successfully, effectively and rapidly detect Xcc race 6 from other Xcc races/strains/pathovars as well as other plant pathogenic bacteria. This is the first report for race-specific molecular markers for Xcc race 6.