Abstract:
:BACKGROUND: A rapid microtiter plate based sandwich hybridization assay was developed for detection and quantification of single RNA species using magnetic beads. Following solution hybridization target RNA molecules were collected by biotin-streptavidin affinity binding and detected by fluorescence signal generated by alkaline phosphatase. The 18S rRNA and SUC2 mRNA of Saccharomyces cerevisiae were used as model RNA target molecules. RESULTS: The sensitivity of the assay was approximately 1.2 x 109 (2 fmol) molecules of target RNA. The developed method was feasible with crude cell lysates of S. cerevisiae carlsbergensis and was evaluated by measuring the levels of 18S rRNA during cell growth and SUC2 mRNA under repressive and inductive conditions. The 18S rRNA expression level followed the changes in the specific growth rate. SUC2 mRNA levels were in good correlation with the measured invertase enzyme activities. CONCLUSIONS: The here presented sandwich hybridisation method was succefully applied for monitoring the amounts of ribosomal RNA and mRNA with high expression level in shake flask cultivation conditions. Sandwich hybridisation method offers a fast and convenient tool for following single key RNA species of interest in the production conditions.
journal_name
Microb Cell Factjournal_title
Microbial cell factoriesauthors
Rautio J,Barken KB,Lahdenperä J,Breitenstein A,Molin S,Neubauer Pdoi
10.1186/1475-2859-2-4keywords:
subject
Has Abstractpub_date
2003-04-28 00:00:00pages
4issue
1issn
1475-2859journal_volume
2pub_type
杂志文章abstract:BACKGROUND:Streptomyces venezuelae ATCC 10712 produces antibiotics chloramphenicol (Cml) and jadomycin (Jad) in response to nutrient limitation and ethanol shock (ES), respectively. Biosynthesis of Cml and Jad was shown to be reciprocally regulated via the action of regulatory proteins JadR1 and JadR2 encoded by the ja...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0484-9
更新日期:2016-05-20 00:00:00
abstract:BACKGROUND:In the biopharmaceutical industry, Escherichia coli (E. coli) strains are among the most frequently used bacterial hosts for producing recombinant proteins because they allow a simple process set-up and they are Food and Drug Administration (FDA)-approved for human applications. Widespread use of E. coli in ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-12-58
更新日期:2013-06-11 00:00:00
abstract:BACKGROUND:The efficiency of biotechnological production processes depends on selecting the best performing microbial strain and the optimal cultivation conditions. Thus, many experiments have to be conducted, which conflicts with the demand to speed up drug development processes. Consequently, there is a great need fo...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-9-86
更新日期:2010-11-13 00:00:00
abstract:BACKGROUND:Oil palm empty fruit bunch (EFB) is a lignocellulosic waste produced in palm oil industry. EFB mainly consists of cellulose, hemicellulose (mainly xylan) and lignin and has a great potential to be reused. Converting EFB to fermentable sugars and value-added chemicals is a much better choice than treating EFB...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-014-0157-5
更新日期:2014-11-11 00:00:00
abstract:BACKGROUND:Numerous challenges remain to achieve industrially competitive space-time yields for bio-oxidations. The ability to rapidly screen bioconversion reactions for characterization and optimization is of major importance in bioprocess development and biocatalyst selection; studies at conventional lab scale are ti...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-017-0788-4
更新日期:2017-10-10 00:00:00
abstract:BACKGROUND:p-Hydroxycinnamic acid (pHCA) is an aromatic compound that serves as a starting material for the production of many commercially valuable chemicals, such as fragrances and pharmaceuticals, and is also used in the synthesis of thermostable polymers. However, chemical synthesis of pHCA is both costly and harmf...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-12-45
更新日期:2013-05-07 00:00:00
abstract:BACKGROUND:Yeast viability and vitality are essential for different industrial processes where the yeast Saccharomyces cerevisiae is used as a biotechnological tool. Therefore, the decline of yeast biological functions during aging may compromise their successful biotechnological use. Life span is controlled by a varie...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-12-1
更新日期:2013-01-02 00:00:00
abstract:BACKGROUND:Recent years have seen a huge growth in the market of industrial yeasts with the need for strains affording better performance or to be used in new applications. Stress tolerance of commercial Saccharomyces cerevisiae yeasts is, without doubt, a trait that needs improving. Such trait is, however, complex, an...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-9-56
更新日期:2010-07-13 00:00:00
abstract:BACKGROUND:Genetic factors and a dysregulated immune response towards commensal bacteria contribute to the pathogenesis of Inflammatory Bowel Disease (IBD). Animal models demonstrated that the normal intestinal flora is crucial for the development of intestinal inflammation. However, due to the complexity of the intest...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-6-18
更新日期:2007-06-06 00:00:00
abstract:: Different species of microorganisms including yeasts, filamentous fungi and bacteria have been used in the past 25 years for the controlled production of foreign proteins of scientific, pharmacological or industrial interest. A major obstacle for protein production processes and a limit to overall success has been th...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-7-11
更新日期:2008-04-04 00:00:00
abstract:BACKGROUND:Terpenoids constitute a large family of natural products, attracting commercial interest for a variety of uses as flavours, fragrances, drugs and alternative fuels. Saccharomyces cerevisiae offers a versatile cell factory, as the precursors of terpenoid biosynthesis are naturally synthesized by the sterol bi...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-10-4
更新日期:2011-01-28 00:00:00
abstract:BACKGROUND:Intracellular metabolism of glucocorticoid hormones plays an important role in the pathogenesis of metabolic syndrome and regulates, among many physiological processes, collagen metabolism in skin. At the peripheral level the concentration of active glucocorticoids is mainly regulated by the 11β-hydroxystero...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0450-6
更新日期:2016-03-15 00:00:00
abstract:BACKGROUND:Lactones are highly valuable cyclic esters of hydroxy fatty acids that find application as pure fragrances or as building blocks of speciality chemicals. While chemical synthesis often leads to undesired racemic mixtures, microbial production allows obtaining optically pure lactones. The production of a spec...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-019-1113-1
更新日期:2019-03-28 00:00:00
abstract:BACKGROUND:Recombinant protein production in the yeast Pichia pastoris is usually based on the alcohol oxidase promoters pAOX1 and pAOX2, which are regulated by methanol and strongly repressed by other C-sources, like glycerol and glucose. However, the use of methanol brings several disadvantages, which is why current ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-015-0292-7
更新日期:2015-07-10 00:00:00
abstract:BACKGROUND:Calreticulin (CRT) resides in the endoplasmic reticulum (ER) and functions to chaperone proteins, ensuring proper folding, and intracellular Ca(2+) homeostasis. Emerging evidence shows that CRT is a multifunctional protein with significant roles in physiological and pathological processes with presence both ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-015-0356-8
更新日期:2015-10-15 00:00:00
abstract:BACKGROUND:Bioethanol produced by the yeast Saccharomyces cerevisiae is currently one of the most promising alternatives to conventional transport fuels. Lignocellulosic hemicelluloses obtained after hydrothermal pretreatment are important feedstock for bioethanol production. However, hemicellulosic materials cannot be...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-014-0145-9
更新日期:2014-10-12 00:00:00
abstract:BACKGROUND:Prions were first identified as infectious proteins associated with fatal brain diseases in mammals. However, fungal prions behave as epigenetic regulators that can alter a range of cellular processes. These proteins propagate as self-perpetuating amyloid aggregates being an example of structural inheritance...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-11-89
更新日期:2012-06-25 00:00:00
abstract:BACKGROUND:Molecular mechanisms generating genetic variation provide the basis for evolution and long-term survival of a population in a changing environment. In stable, laboratory conditions, the variation-generating mechanisms are dispensable, as there is limited need for the cell to adapt to adverse conditions. In f...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-11-11
更新日期:2012-01-20 00:00:00
abstract::Recent evidence has demonstrated that cell-to-cell signaling is a fundamental activity carried out by numerous microorganisms. A number of specialized processes are reported to be regulated by density-dependent signaling molecules including antibiotic production, bioluminescence, biofilm formation, genetic competence,...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-1-5
更新日期:2002-12-20 00:00:00
abstract:BACKGROUND:In past years research has focused on the development of alternative Gram positive bacterial expression systems to produce industrially relevant proteins. Brevibacillus choshinensis is an easy to handle non-sporulating bacterium, lacking extracellular proteases, that has been already shown to provide a high ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-12-12
更新日期:2013-02-01 00:00:00
abstract::Pseudomonas sp. JHN decolourized and biotransformed 4-chloro-2-nitrophenol (4C2NP) in the presence of additional carbon source. The effect of the various concentrations of the 4C2NP was studied on the decolourization of 4C2NP by Pseudomonas sp. JHN. It was observed that strain JHN decolourized and biotransformed 4C2NP...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-014-0110-7
更新日期:2014-08-12 00:00:00
abstract:BACKGROUND:Probiotic bacteria are increasingly used as immunomodulatory agents. Yet detailed molecular knowledge on the immunomodulatory molecules of these bacteria is lagging behind. Lipoteichoic acid (LTA) is considered a major microbe-associated molecular pattern (MAMP) of Gram-positive bacteria. However, many detai...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-11-161
更新日期:2012-12-15 00:00:00
abstract:BACKGROUND:Co-expression of two distinct guide RNAs (gRNAs) has been used to facilitate the application of CRISPR/Cas9 system in fields such as large genomic deletion. The paired gRNAs are often placed adjacently in the same direction and expressed individually by two identical promoters, constituting direct repeats (D...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-020-01321-4
更新日期:2020-03-10 00:00:00
abstract:BACKGROUND:Microalgae has been concerned as a potential source of biodiesel in the recent years. However, it is costly to harvest microalgae as it is commonly cultured in water and the cells are too small to harvest. In order to reduce the cost of cultivation and harvesting, it is important to improve the biomass produ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-014-0142-z
更新日期:2014-10-01 00:00:00
abstract::Nanotechnology is increasingly using both materials and nano-objects synthesized by living beings, most of them produced by microbial cells. Emerging technologies and highly integrative approaches (such as 'omics and systems biology), that have been largely proven successful for the production of proteins and secondar...
journal_title:Microbial cell factories
pub_type: 社论
doi:10.1186/1475-2859-9-53
更新日期:2010-07-05 00:00:00
abstract:BACKGROUND:Crocin is a carotenoid-derived natural product found in the stigma of Crocus spp., which has great potential in medicine, food and cosmetics. In recent years, microbial production of crocin has drawn increasing attention, but there were no reports of successful implementation. Escherichia coli has been engin...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-019-1166-1
更新日期:2019-07-05 00:00:00
abstract:BACKGROUND:Bacterial surface display is of interest in many applications, including live vaccine development, screening of protein libraries and the development of whole cell biocatalysts. The goal of this work was to understand which parameters result in production of large quantities of cells that at the same time ex...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-10-72
更新日期:2011-09-14 00:00:00
abstract:BACKGROUND:The Bacillus subtilis spore has long been used to display antigens and enzymes. Spore display can be accomplished by a recombinant and a non-recombinant approach, with the latter proved more efficient than the recombinant one. We used the non-recombinant approach to independently adsorb two thermophilic enzy...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-017-0833-3
更新日期:2017-11-28 00:00:00
abstract:BACKGROUND:Fine-tuning the aeration for cultivations when oxygen-limited conditions are demanded (such as the production of vaccines, isobutanol, 2-3 butanediol, acetone, and bioethanol) is still a challenge in the area of bioreactor automation and advanced control. In this work, an innovative control strategy based on...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-019-1198-6
更新日期:2019-09-04 00:00:00
abstract:BACKGROUND:E. coli is a robust host for various genetic manipulations and has been used commonly for bioconversion of hexose and pentose sugars into valuable products. One of the products that E. coli make under fermentative condition is ethanol. However, availability of limited reducing equivalence and generation of c...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-11-145
更新日期:2012-11-04 00:00:00