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Expression of Tissue Inhibitor of Metalloproteinases by Recombinant Baculovirus-Infected Insect Cells Cultured in an Airlift Fermentor
Time of Update: 2021-02-22
In this system, the gene to be expressed is cloned into a plasmid transfer vector, downstream of a strong baculovirus gene promoter, and cotransfected with wildtype virusDNA into insect cells.
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DNA Extraction from Natural Environments
Time of Update: 2021-02-22
Until recently, studies on microbial communities present in natural environments relied on conventional optical microscopic observation and cultivation-based approaches.
The most commonly cited limitation stems from the finding that the majority of microbial cells in natural environments cannot be cultured in the laboratory—the phenomenon of nonculturability.
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Insect Cell- Culture Techniques in Serum- Containing Medium
Time of Update: 2021-02-22
The establishment of new lines is now routine, and the culture of hundreds of cell lines from a variety of insect species have to date been reported ( 2 ,4 ,5 ,6 ).
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Detection and Differentiation of Ruminant Mycoplasmas
Time of Update: 2021-02-22
While a considerable part of this group is conceived to be of minor epidemiological relevance, it contains some important pathogenic agents that have specific host ranges.
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Molecular Detection of Drug-Resistant Mycobacterium tuberculosis with a Scanning-Frame Oligonucleotide Microarray
Time of Update: 2021-02-22
In this chapter, we describe the utility of a microarray-based approach exploiting short overlapping oligonucleotides (sliding-frame array) to rapidly detect drug resistance–associated mutations (substitutions, deletions, and insertions) in thepncA gene responsible for resistance of M.
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Virus-like Particle and Viral Vector Production Using the Baculovirus Expression Vector System/Insect Cell System: Adeno-Associated Virus-Based Products
Time of Update: 2021-02-22
The ability to make a large variety of virus-like particles (VLPs) has been successfully achieved in the baculovirus expression vector system (BEVS)/insect cell system.
This chapter focuses on the methodology, based on the work done in our lab, for the production of AAV-like particles and vectors in a BEVS/insect cell system.
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Scanning Low-Frequency Point Mutants in the Mitochondrial Genome Using Constant Denaturant Capillary Electrophoresis
Time of Update: 2021-02-22
We have developed a highly sensitive technique termedconstant denaturant capillary electrophoresis (CDCE) ( 1 ), permitting measurement of mitochondrial point mutations at fractions as low as 10 6 in human cells and tissues ( 2 ).
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Morphometry of Rectal Mucosa
Time of Update: 2021-02-22
Morphometry was first introduced into pathology 80 yr ago.
Thus, in comparative morphometric studies, it is essential to ensure that only sections obtained in proper and consistent orientation are used ( 2 , 7 , 8 ).
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Evaluation of Rotavirus Vaccines in Small Animal Models
Time of Update: 2021-02-22
Because RV is a localized enteric infection, and induction of intestinal mucosal immune responses was expected to be required for protection, live orally administered vaccines were pursued first.
The nonreplicating immunogen must be able to induce protective immune responses against the target virus.
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Monitoring Microbial Activities Using Membrane Inlet Mass Spectrometry
Time of Update: 2021-02-22
Currently used methods include manometric techniques, microsensors, chemical assays, gas chromatography and high-performance liquid chromatography, but all have their limitations and usually require substantial disruption to the environment being studied.
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Immunological Methods for the Detection of Human Cytomegalovirus
Time of Update: 2021-02-22
In addition, HCMV has been implicated as a co-etiological agent in cervical cancer ( 2 ) and has been found associated with a wide range of other tumors ( 1 ).
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Analysis of Chromatin Organization by Deep Sequencing Technologies
Time of Update: 2021-02-22
Protection from MNase cleavage has often been used to map transcription factor binding sites and nucleosomal positions on a single-gene basis; however, by combining MNase digestion with high-�throughput, paired-end DNA sequencing, it is now possible to simultaneously map DNA-protein interaction regions across the entire genome.
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Biosynthesis, Extraction, Purification, and Analysis of Trisporoid Sexual Communication Compounds from Mated Cultures of Blakeslea trispora
Time of Update: 2021-02-22
The zygomyceteBlakeslea trispora produces high amounts of the general zygomycete β-carotene-derived sexual signal compounds, the trisporoids.
The effect of the extraction and activity of the isolated compounds is best tested physiologically, exploiting the ability of trisporoids to induce the formation of sexually committed hyphae, the zygophores, in other zygomycete species.
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Gene Replacement in Mycobacterium intracellulare
Time of Update: 2021-02-22
Much of the present knowledge of the bacterial cell emanates from studies of spontaneous mutants and mutants obtained as a result of random or directed mutagenesis.
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Extraction of Yeast Lipids
Time of Update: 2021-02-22
Quantitative extraction of lipids from the tissue or microorganism of choice is key to their subsequent analysis. In this chapter, we describe a simple and rapid protocol that relies on glass bead di
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Preparation of Chromatin Assembly Extracts from Xenopus Oocytes
Time of Update: 2021-02-22
The majority ofDNA in eukaryotic cells is packaged by histones and many poorly characterized nonhistone proteins to form a dynamic structure known as chromatin.
Moreover, histones, via protein-protein interactions or by playing an architectural role, can facilitate or inhibit the transcriptional activation process ( 1 , 2 ).
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Analysis of Chromatin by Scanning Force Microscopy
Time of Update: 2021-02-22
The most widely used probe microscopy for biological applications, the scanning force microscope (SFM), also known as the atomic force microscope, is capable of imaging samples under very mild conditions (for recent reviews,see refs.
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Measuring Oxidative mtDNA Damage and Repair Using Quantitative PCR
Time of Update: 2021-02-22
The human mitochondrial genome was completely sequenced in 1981 by Anderson and co-workers ( 1 ) and consists of a closed circular supercoiledDNA molecule of 16,569 base pairs.
The mitochondrial genome encodes 13 polypeptides, 22 transfer RNAs (tRNAs), and 2 rRNA.
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Preparation of Culture Filtrate Proteins from Mycobacterium tuberculosis
Time of Update: 2021-02-22
Culture filtrates obtained by in vitro cultivation ofMycobacterium tuberculosis have been studied for more than 20 years to identify and characterize proteins of immunological relevance.
Culture filtrate preparations have been shown to induce a protective immune response in mice and guinea pigs by several groups ( 1 – 4 ).
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Reproducible Quantitative PCR of Mitochondrial and Nuclear DNA Copy Number Using the LightCycler
Time of Update: 2021-02-22
Recent developments inPCR fluorimetry have allowed for quick quantification of target molecules. Before the invention of fluorimetric quantitative PCR, researchers who wanted to quantify the amount
