Kapitel 8 Flashcards
Cloning vector that can accommodate large pieces of DNA - typically up to 1 million base pairs.
bacterial artificial chromosome (BAC)
Clone containing double-stranded cDNA molecules derived from the protein-coding mRNA molecules present in a cell.
cDNA clone
Collection of cloned DNA molecules representing complementary DNA copies of the mRNA produced by a cell.
cDNA library
Name given to a strategy that uses large-scale screening of hundreds of thousands of small molecules in biological assays to identify chemicals that affect a particular biological process and that can then be used to study it.
chemical biology
Technique by which chromosomal DNA bound by a particular protein can be isolated and identified by precipitating it by means of an antibody against the protein. (Figures 8–66 and 8–67)
chromatin immunoprecipitation
Technique for separation of a mixture of substances in solution by passage through a column containing a porous solid matrix. Substances are retarded to different extents by their interaction with the matrix and can be collected separately from the column. Depending on the matrix - separation can be according to charge - hydrophobicity - size - or the ability to bind to other molecules.
column chromatography
Test to determine whether two mutations that produce similar phenotypes are in the same or different genes. (Panel 8–2 - pp. 487)
complementation test
Mutation that changes a protein or RNA molecule so that its function is altered only under some conditions - such as at an unusually high or unusually low temperature.
conditional mutation
See RNA-seq
deep RNA sequencing
The standard enzymatic method of DNA sequencing. (Panel 8–1 - p. 478)
dideoxy sequencing
(1) The act of making many identical copies (typically billions) of a DNA molecule—the amplification of a particular DNA sequence. (2) Also - the isolation of a particular stretch of DNA (often a particular gene) from the rest of the cell’s genome.
DNA cloning
Collection of cloned DNA molecules - representing either an entire genome (genomic library) or complementary DNA copies of the mRNA produced by a cell (cDNA library).
DNA library
A large array of short DNA molecules (each of known sequence) bound to a glass microscope slide or other suitable support. Used to monitor expression of thousands of genes simultaneously: mRNA isolated from test cells is converted to cDNA - which in turn is hybridized to the microarray. (Figure 8–64)
DNA microarray
Analysis to discover the order in which the genes act - by investigating if a mutation in one gene can mask the effect of a mutation in another gene when both mutations are present in the same organism or cell.
epistasis analysis
Engineered protein that combines two or more normally separate polypeptides. Produced from a recombinant gene.
fusion protein
Procedure for discovery of genes affecting a specific phenotype by surveying large numbers of mutagenized individuals.
genetic screen
The study of the genes of an organism on the basis of heredity and variation.
genetics
Process attempting to mark out all the genes (protein-coding and noncoding) in a genome and ascribing functions to each.
genome annotation
Collection of cloned DNA molecules representing an entire genome.
genomic library
Genetic constitution of an individual cell or organism. The particular combination of alleles found in a specific individual. (Panel 8–2 - p. 486)
genotype
Combination of alleles and DNA markers that has been inherited in a large - linked block on one chromosome of a homologous pair—undisturbed by genetic recombination—across many generations.
haplotype block
Type of chromatography that uses columns packed with tiny beads of matrix; the solution to be separated is pushed through under high pressure.
high-performance liquid chromatography (HPLC)
Hybrid cell line generated by fusion of a tumor cell and another cell type. Monoclonal antibodies are produced by hybridoma lines obtained by fusing antibody-secreting B cells with cells of a B lymphocyte tumor. (Figure 8–4)
hybridoma
See Western blotting
immunoblotting
Antibody secreted by a hybridoma cell line. Because the hybridoma is generated by the fusion of a single B cell with a single tumor cell - each hybridoma produces antibodies that are all identical. (Page 444)
monoclonal antibody
NMR is the resonant absorption of electromagnetic radiation at a specific frequency by atomic nuclei in a magnetic field - due to flipping of the orientation of their magnetic dipole moments. The NMR spectrum provides information about the chemical environment of the nuclei. NMR is used widely to determine the three-dimensional structure of small proteins and other small molecules. The principles of NMR are also used for medical diagnostic purposes in magnetic resonance imaging (MRI). (Figure 8–22)
nuclear magnetic resonance (NMR) spectroscopy
A continuous nucleotide sequence free from stop codons in at least one of the three reading frames (and thus with the potential to code for protein).
open reading frame (ORF)
The observable character (including both physical appearance and behavior) of a cell or organism. (Panel 8–2 - p. 486)
phenotype
Small - circular molecules of double-stranded DNA derived from plasmids that occur naturally in bacterial cells; widely used for gene cloning.
plasmid vector
Technique for amplifying specific regions of DNA by the use of sequence-specific primers and multiple cycles of DNA synthesis - each cycle being followed by a brief heat treatment to separate complementary strands. (Figure 8–36)
polymerase chain reaction (PCR)
Describes genome sequences that coexist as two or more sequence variants at high frequency in a population.
polymorphisms
Fractionated cell homogenate that retains a particular biological function of the intact cell - and in which biochemical reactions and cell processes can be more easily studied.
purified cell-free system
Technique in which a population of mRNAs is converted into cDNAs via reverse transcription - and the cDNAs are then amplified by PCR. The quantitative part relies on a direct relationship between the rate at which the PCR product is generated and the original concentration of the mRNA species of interest.
quantitative RT-PCR (reverse transcription–polymerase chain reaction
Collection of techniques by which DNA segments from different sources are combined to make a new DNA - often called a recombinant DNA. Recombinant DNAs are widely used in the cloning of genes - in the genetic modification of organisms - and in the production of large amounts of rare proteins.
recombinant DNA technology
One of a large number of nucleases that can cleave a DNA molecule at any site where a specific short sequence of nucleotides occurs. Extensively used in recombinant DNA technology. (Figure 8–24)
restriction nuclease
Approach to discovering gene function that starts from the DNA (gene) and its protein product and then creates mutants to analyze the gene’s function.
reverse genetics
Sequencing the entire repertoire of RNA from a cell or tissue; also known as deep RNA sequencing.
RNA-seq
The ability of biological regulatory systems to function normally in the face of perturbations such as exposure to frequent and/or extreme variations in external conditions or the concentrations or activities of key components.
robustness
See dideoxy sequencing
Sanger sequencing
Type of electrophoresis used to separate proteins by size. The protein mixture to be separated is first treated with a powerful negatively charged detergent (SDS) and with a reducing agent (β-mercaptoethanol) - before being run through a polyacrylamide gel. The detergent and reducing agent unfold the proteins - free them from association with other molecules - and separate the polypeptide subunits.
sodium dodecyl sulfate polyacrylamide-gel electrophoresis (SDS-PAGE)
Random. Involving chance - probability - or random variables.
stochastic
The foreign or modified gene that has been added to create a transgenic organism.
transgene
Plant or animal that has stably incorporated one or more genes from another cell or organism (through insertion - deletion - and/or replacement) and can pass them on to successive generations. (Figures 8–53 and 8–70)
transgenic organism
Technique combining two different separation procedures—separation by charge (isoelectric focusing) in the first dimension - then separation by size in a direction at a right angle to that of the first step—to resolve up to 2000
two-dimensional gel electrophoresis
Technique by which proteins are separated by electrophoresis and immobilized on a paper sheet and then analyzed - usually by means of a labeled antibody. Also called immunoblotting.
Western blotting
Technique for determining the three-dimensional arrangement of atoms in a molecule based on the diffraction pattern of x-rays passing through a crystal of the molecule. (Figure 8–21)
x-ray crystallography
