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| Registros recuperados: 978 | |
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| Bazzini,Ariel A; Mongelli,Vanesa C; Hopp,H. Esteban; del Vas,Mariana; Asurmendi,Sebastián. |
| Gene silencing, also called RNA interference (RNAi) is a specific mechanism of RNA degradation involved in gene regulation, development and defense in eukaryotic organisms. It became an important subject in the teaching programs of molecular biology, genetics and biotechnology courses in the last years. The aim of this work is to provide simple and inexpensive assays to understand and teach gene silencing using plants as model systems. The use of transient and permanent transgenic plants for expressing reporter genes, like those derived from jellyfish green fluorescent protein (gfp) encoding gene, provides a nice, colorful and conclusive image of gene silencing. Three experimental approaches to evidence RNA silencing are depicted. In the first approach... |
| Tipo: Journal article |
Palavras-chave: Gene silencing; GFP; HC-Pro; PDS; Protocols; PTGS; Teaching; Transgenic; Transient expression; TRV; VIGS. |
| Ano: 2007 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582007000200002 |
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| Mohd. Raih,Mohd. Firdaus; Sailan,Ahmad Tarmidi; Zamrod,Zulkeflie; Embi,Mohd. Noor; Mohamed,Rahmah. |
| Cytochrome c oxidase, the terminal enzyme of the respiratory chains of mitochondria and aerobic bacteria, catalyzes electron transfer from cytochrome c to molecular oxygen. The enzyme belongs to the haem-copper-containing oxidases superfamily. A recombinant plasmid carrying a 2.0 kb insert from a Burkholderia pseudomallei genomic library was subjected to automated DNA sequencing utilizing a primer walking strategy. Analysis of the 2002 bp insert revealed a 1536 bp open reading frame predicted to encode a putative cytochrome c oxidase. Further analysis using sequence alignments and tertiary structure analysis tools demonstrated that the hypothetical B. pseudomallei cytochrome c oxidase is similar to cytochrome c oxidases from other organisms such as Thermus... |
| Tipo: Journal article |
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| Ano: 2003 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582003000100005 |
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| Cui,Yihong; Zhu,Guangqin; Chen,Qiuju; Wang,Yunfei; Yang,Mingming; Song,Yuxuan; Wang,Jiangang; Cao,Binyun. |
| Complementary DNA (cDNA) is valuable for investigating protein structure and function in the research of life science, but it is difficult to obtain by traditional reverse transcription. In this study, we employed a novel strategy to clone the human leukemia inhibitory factor (hLIF) gene cDNA from genomic DNA directly isolated from the mucous membrane of mouth. The hLIF sequence can be acquired within a few hours by means of amplification of each exon and splicing using overlap-PCR. Thus, the new approach developed in this study is simple, time- and cost-effective, and it is not limited to particular gene expression levels of each tissue. |
| Tipo: Journal article |
Palavras-chave: CDNA cloning; HLIF; Oral mucous membrane; Overlap PCR. |
| Ano: 2011 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000300012 |
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| Kuhn,Jennifer; Müller,Hagen; Salzig,Denise; Czermak,Peter. |
| Background The purpose of this work was to find a rapid method for glycerol detection during microbial fermentations. The method requirements were, first, to avoid sample pretreatment, and second, to measure glycerol precisely especially out of fermentation broth. Results This was achieved by combining two reaction principles - the Malaprade reaction and the Hantzsch reaction. In the Malaprade reaction, glycerol is converted into formaldehyde. This forms a dye in the Hantzsch reaction after which adsorption is than detected. The subsequent assay was investigated with two different fermentation media, a chemically undefined and a chemically defined media, used for Pichia pastoris fermentation. In both media, as well as in real fermentation samples, glycerol... |
| Tipo: Journal article |
Palavras-chave: Glycerol determination; Fermentation broth; Hantzsch reaction; Malaprade reaction. |
| Ano: 2015 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582015000300017 |
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| Chen,Sique; Zheng,Xiujuan; Cao,Hongrui; Jiang,Linghui; Liu,Fangqian; Sun,Xinli. |
| Background Thermostable DNA polymerase (Taq Pol ?) from Thermus aquaticus has been widely used in PCR, which was usually extracted with Pluthero's method. The method used ammonium sulfate to precipitate the enzyme, and it saved effort and money but not time. Moreover, we found that 30-40% activity of Taq Pol I was lost at the ammonium sulfate precipitation step, and the product contained a small amount of DNA. Results We provided a novel, simplified and low-cost method to purify the Taq Pol ? after overproduction of the enzyme in Escherichia coli, which used ethanol instead of ammonium sulfate to precipitate the enzyme. The precipitate can be directly dissolved in the storage buffer without dialysis. In addition, DNA and RNA contamination was removed with... |
| Tipo: Journal article |
Palavras-chave: Ethanol precipitation; PCR; Purification; Taq DNA polymerase. |
| Ano: 2015 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582015000500005 |
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| Zhao,Yongliang; Dong,Zongming; Li,Tengfei; Huang,Gang. |
| Background: The AdEasy system is a fast-track system for generating recombinant adenoviruses using the efficient homologous recombination machinery between shuttle and adenovirus backbone plasmids in Escherichia coli BJ5183 cells. The key step is homologous recombination in BJ5183 cells, which is driven by RecA activity. However, culture time is stringently limited to reduce the damage to recombinant plasmids by RecA activity. Therefore, rapid identification of recombinant adenoviruses within the limited time-period is critical. Results: We developed a simple negative selection method to identify recombinant adenoviruses using colony PCR, which improves the efficiency of adenovirus recombination screening and packaging. Conclusions: The negative selection... |
| Tipo: Journal article |
Palavras-chave: Adenovirus; AdEasy; Colony PCR; Homologous recombination; RecA. |
| Ano: 2014 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582014000100008 |
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| Sarwat,Maryam; Singh Negi,Madan; Lakshmikumaran,Malathi; Kumar Tyagi,Akhilesh; Das,Sandip; Shankar Srivastava,Prem. |
| For studying genetic diversity in natural populations of Terminalia, a medicinal plant, our attempts to isolate high quality DNA using several previously reported protocols and even modifications were unsuccessful. We therefore combined CTAB based isolation, and column based purification step, to isolate DNA from Terminalia arjuna. The DNA isolated using this standardized protocol was high in quality and suitable for restriction digestion and generation of random amplification of polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP). |
| Tipo: Journal article |
Palavras-chave: DNA extraction; Medicinal plants; Molecular markers; Terminalia species. |
| Ano: 2006 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000100011 |
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| Li,Biaoru. |
| Recent advances in functional genomics allow us to estimate the expression of several thousands of genes in the mammalian genome. Techniques such as microarrays, expressed tag sequencing (EST), serial analysis of gene expression (SAGE), subtractive cloning and differential display (DD), and two-dimensional electrophoresis gel have been extensively used to screen and analyze parallel gene expression. Some pathological processes, for example, tumorigenesis and solid tumour growth, in which the former is derived from a single-cell and the latter has a mixed-cell problem, present new challenges to the limit of these functional genomic techniques. To fully understand the functions of cells in tumorigenesis or in heterogeneous solid tumour masses, it is... |
| Tipo: Journal article |
Palavras-chave: Genome; Genomic expression; Genomic expression analysis at single cell; Single cell. |
| Ano: 2005 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582005000100011 |
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| Wang,Zhejian; Ma,Zhao; Wang,Lili; Tang,Chengchen; Hu,Zhibi; Chou,GuiXin; Li,Wankui. |
| Background An endophytic fungus lives within a healthy plant during certain stages of, or throughout, its life cycle. Endophytic fungi do not always cause plant disease, and they include fungi that yield different effects, including mutual benefit, and neutral and pathogenic effects. Endophytic fungi promote plant growth, improve the host plant's resistance to biotic and abiotic stresses, and can produce the same or similar biologically active substances as the host. Thus, endophytic fungal products have important implications in drug development. Result Among the numerous endophytic fungi, we identified two strains, L10Q37 and LQ2F02, that have anti-acetylcholinesterase activity, but the active compound was not huperzine A. The aim of this study was to... |
| Tipo: Journal article |
Palavras-chave: Anti-acetylcholinesterase activity; Acetylcholinesterase inhibitor; Biological characteristics; Endophytic fungi. |
| Ano: 2015 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582015000600003 |
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| Felmer,Ricardo; Arias,María Elena. |
| Total number of cells in cloned embryos is generally lower than that of in vivo derived embryos and in bovines cell allocation at the blastocyst stage, has been observed to be affected in a large proportion of cloned embryos. The current embryo staining procedures are toxic for mammalian cells and thus can not be used to determine the developmental potential of a stained embryo. Therefore, in the present study we sought to assess the feasibility to develop a noninvasive embryo model that would be suitable for the evaluation of cloned embryos subjected to different nuclear transfer and embryo culture procedures. For doing this, we stably transfected a bovine embryonic fibroblast cell line and generated a number of clones that constitutively expressed a red... |
| Tipo: Journal article |
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| Ano: 2008 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000500002 |
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| Wang,Ping; Nong,Xu-Hua; Ge,Jie-Hong. |
| Nonylphenol ethoxylates (NPEOs), which are widely used for industrial and domestic purposes, exert adverse effects on wildlife after being used and discharged into the environment. However, their ultimate biodegradability and biodegradation pathway remains unclear. In this study, the aerobic degradability of nonylphenol ethoxylates (NPEOs) by the acclimated microorganisms in active sludge was examined using shaking-flask tests. The degradation of benzene rings in NPEOs was determined using UV spectroscopy and high performance liquid chromatography (HPLC). Results showed that more than 80% of benzene rings were removed after 8-10 days of degradation, and the majority of NPEOs were also removed after 9 days of degradation, indicating NPEOs and the benzene... |
| Tipo: Journal article |
Palavras-chave: Aerobic biodegradation; Degradation pathway; ESI-MS; HPLC; Nonylphenol ethoxylates. |
| Ano: 2011 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000400001 |
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| Registros recuperados: 978 | |
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