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Genetic Modification in Bacillus subtilis for Production of C30 Carotenoids
Time of Update: 2021-02-16
This chapter describes a method to synthesize C30 carotenoids inBacillus subtilis , which is generally recognized as a safe (GRAS) organism.
aureus crtM andcrtN genes intoB.
Thus, carotenoid biosynthesis genes ofS.
subtilis in order to construct a safe organism producing C30 carotenoids.
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High-Level Periplasmic Expression and Purificationof scFvs
Time of Update: 2021-02-16
The isolation of recombinant antibodies by phage display naturally leads to experiments to evaluate their biological and immunological properties. Although crude preparations may have their value in
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Analysis of the Efficiency of Adenovirus Transcription
Time of Update: 2021-02-16
This method is designed to measure rates of transcription from adenoviral promoters as a function of the concentrations within infected cells of the promoter(s) of interest.
The latter parameter is assessed by quantification of viralDNA by hybridization of membrane-bound DNA following purification of DNA from nuclear fractions of adenovirus-infected cells.
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Flow Cytometry in Environmental Microbiology: A Rapid Approach for the Isolation of Single Cells for Advanced Molecular Biology Analysis
Time of Update: 2021-02-16
The isolation and subsequent characterization of microbial cells from within environmental samples is a difficult process.
We describe here the combination of fluorescent in situ hybridization and cell sorting for the detection and isolation of Candidate Division TM7 bacteria from an enriched soil sample.
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Identification of Protein Complexes from Filamentous Fungi with Tandem Affinity Purification
Time of Update: 2021-02-16
Tandem affinity purification (TAP) allows the enrichment of native protein complexes from cell extracts under mild conditions.
We codon-optimized tags and established TAP, previously not applicable to filamentous fungi, for the model organismAspergillus nidulans.
nidulans which can also be adapted to other filamentous fungi.
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Two Efficient Methods for the Conjugation of Smooth-Form Lipopolysaccharides with Probes Bearing Hydrazine or Amino Groups. I. LPS Activation with Cyanogen Bromide
Time of Update: 2021-02-16
This chapter presents a conjugation method for coupling probes bearing hydrazine or primary amino groups to a smooth(S)-form lipopolysaccharide (LPS).
Minnesota with dansyl hydrazine and horseradish �peroxidase yields labeling ratios above 300 nmol dansyl per mg LPS, with nearly no loss of the original endotoxin activity.
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Ribozyme-Mediated Inhibition of Cell Proliferation: A Model for Identifying and Refining a Therapeutic Ribozyme
Time of Update: 2021-02-16
The sequence specificity is determined by the ability of the ribozyme to base pair with nucleotides near the cleavage site of the target RNA; by altering the substrate recognition sequences, intramolecular,cis -cleaving ribozymes can be engineered to cleave intrans (reviewed in Cech, T.
In theory, ribozymes can be engineered to cleave any RNA species, site-specifically, in the background of cellular RNA.
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Purification of the Extracellular Domain of the Epidermal Growth Factor Receptor Produced by Recombinant Baculovirus-Infected Insect Cells in a 10-L Reactor
Time of Update: 2021-02-16
The binding of ligand to the extracellular domain of the epidermal growth factor (EGF) receptor is the event that initiates the activation of the tyrosine kinase activity present on the cytoplasmic domain.
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Medical Significance of Mycoplasmas
Time of Update: 2021-02-16
Other mycoplasmas have been isolated from predominantly respiratory and genito-urinary sites.
Table 1 Mycoplasmas Isolated from Humans LocationOccurrenceSpeciesO, oropharynx; R, respiratory;GU, genitourinary+ Rare,++ Common,+++ Widespread M.
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Detection of Clostridium botulinum by Multiplex PCR in Foods and Feces
Time of Update: 2021-02-16
Clostridium botulinum is a diverse group of anaerobic spore-forming organisms that produce lethal botulinum neurotoxins (BoNT) during their growth.
The most common forms of human botulism include the classical foodborne botulism due to ingestion of BoNT preformed in food, and infant botulism due to spore germination, growth, and toxin production in the infant’s intestine.
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Selectable Markers for Eukaryotic Cells
Time of Update: 2021-02-16
The transfer of DNA sequences into a population of cells can rarely, if ever, be achieved with 100% efficiency.
If the transferred sequences do not confer a selective growth advantage, it is essential to use selection for transduced cells.
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Using an Advanced Microfermentor System for Strain Screening and Fermentation Optimization
Time of Update: 2021-02-16
To maximize the bioproduction of a selected strain, bioreaction (fermentation) conditions need to be optimized.
Bench-scale fermentor experiments (>0.5 L) are well controlled under designed conditions and provide high-quality data, but they are also very time- and cost-consuming.
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Electronic Access Mycobacterium tuberculosis Sequence Data
Time of Update: 2021-02-16
This information is obviously of great benefit to researchers in the field, but the sheer volume of data often makes it inaccessible.
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PCR-Derived Methods for the Identification of Trichinella Parasites from Animal and Human Samples
Time of Update: 2021-02-16
Trichinella worms (family: Trichinellidae; phylum: Nematoda) are parasites that mainly infect mammals, including humans, although they have been found in birds and, recently, in African crocodiles ( 1 , 2 ).
The main reservoir is represented by carnivores with cannibalistic and scavenger behavior.
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Rapid Identification of Mycobacteria and Rapid Detection of Drug Resistance in Mycobacterium tuberculosis in Cultured Isolates and in Respiratory Specimens
Time of Update: 2021-02-16
In this chapter, several simple nucleic acid amplification-based techniques are introduced as molecular approach for clinical diagnosis of tuberculosis.
Despite the high specificity and sensitivity of the molecular techniques, mycobacterial culture remains the “Gold Standard” for tuberculosis diagnosis.
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Pyrolysis Mass Spectrometry
Time of Update: 2021-02-16
Pyrolysis of microorganisms yields complex mixtures of products that can be analyzed quantitatively by mass spectrometry (MS). The resulting mass spectra represent transient bacterial “fingerprints”
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Electroporation of Francisella tularensis
Time of Update: 2021-02-16
In North America, tularemia presents as an acute febrile lymphadenitis that can progress into a life-threatening pneumonic illness ( 1 ).
In Europe and Asia, tularemia is often acquired from consumption of contaminated water or inhalation of contaminated dust.
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Origins and Development of Peptide Antibiotic Research: From Extracts to Abstracts to Contracts
Time of Update: 2021-02-16
Peptide antibiotic research, which in the larger sense includes protein antibiotic research, actually began during the late 19th century with the work of Ehrlich, Metchnikov, Kanthack, and Petterson.
He noted that the granules of approx 2 or 3% of the cells stained intensely with eosin, an acid dye.
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Expression of Foreign Genes in Bombyx mori Larvae Using Baculovirus Vectors
Time of Update: 2021-02-16
Although both the AcNPV and BmNPV systems provide highlevel expression of foreign genes using larval hosts ( 3 ,4 ), the silkworm larva offers several additional advantages, i.e., it is easy to rear, it is large and easy to manipulate, it has a relatively short life cycle (approx 7 wk), and its genetics and biology have been well documented.
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Current energy
Time of Update: 2021-02-16
The so-called current mainly refers to the more stable flow of seafloor water and the channel and the regular flow of sea water due to tides.
Generally speaking, the current energy of a water channel with a maximum flow rate of 2m/s or more has the value of actual development.
