遗传学实验技术

A large number of processes within the cell, in particular the regulation of gene expression, rely on the binding of proteins to specific sites on the DNA. A primary ingredient to understanding these processes is the characterization of the protein-DNA interaction (1). Such a characterizati ...

Polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) (1) is a simple method that allows one to rapidly determine whether there are sequence differences between relatively short stretches of DNA. Coupled with sequence analysis, SSCP is an extremely useful ...

First described by Newton and colleagues in 1989 (1), amplification refractory mutations system (ARMS) has become a standard technique that allows the discrimination of alleles that differ by as little as 1 bp. The system is simple, reliable, and nonisotopic. It clearly distinguishes hete ...

Genetic analysis by restriction fragment length polymorphism (RFLP) is one of the most common methods used to examine nucleic acids for the presence of known sequence variants. A segment that is to be searched for a mutation is amplified from genomic DNA or cDNA, digested by the appropriate restr ...

The amount of genomic DNA available for genetic studies can often be limiting. Degenerated oligonucleotide-primed polymerase chain reaction (DOP-PCR) is an appropriate method for overcoming these limitations by efficiently performing whole genome amplification. The DOP-P ...

An improved nonradioactive polymerase chain reaction (PCR)-based method for simultaneous amplification of multiple loci of microsatellites has been developed as a rapid way to screen for microsatellites (1). The approach, termed multiplex-touchdown PCR (MT-PCR), is performed ...

Repetitive sequences like short tandem repeat (STR) loci are generally referred to as slippery DNA (1). They owe this nickname to a characteristic leading to slippage within the primer-template complex during PCR elongation of the new strand (2,3), resulting in the synthesis of byproducts s ...

Microsatellites are widely distributed highly repetitive DNA sequences composed of di-, tri-, or tetranucleotide repeats (1). They are spread over the whole human genome and are located between and within genes. Physiologically, they exhibit high levels of polymorphism, relative to d ...

RNA editing within the mitochondria of kinetoplastid protozoa is performed by a multicomponent �macromolecular machine known as the editosome. Editosomes are high molecular mass protein assemblies that consist of about 15–25 individual polypeptides. They bind pre-edited tra ...

In vivo mutational analysis is often required to characterize enzymes that function as subunits of the U-insertion/deletion RNA editing core complex (RECC) in mitochondria of Trypanosoma brucei. The mutations may skew phenotypic manifestation of a dominant negative overexpres ...

Evidence for the chemical conversion of adenosine-to-inosine (A-to-I) in messenger RNA (mRNA) has been detected in numerous metazoans, especially those “most successful” phyla: Arthropoda, Mollusca, and Chordata. The requisite enzymes for A-to-I editing, ADARs (adenosine deami ...

Glutamate is the major excitatory neurotransmitter in the mammalian nervous system. The properties of their ionotropic glutamate receptors largely determine how different neurons respond to glutamate. RNA editing in pre-mRNAs encoding subunits of glutamate receptors, part ...

Adenosine-to-inosine (A-to-I) RNA editing is a biologically important posttranscriptional processing event involved in the transcriptome diversification. The most conventional method of editing site identification is to compare the cDNA sequence with its corresponding ...

Many agents that possess antitumor activity have been shown to bind to DNA. Several clinically used chemotherapeutic drugs are alkylating agents, which are known to bind covalently to, and in many cases crosslink, the bases of DNA. These include the nitrogen mustard, chloroethyl-nitroso ...

A large category of anticancer drugs owe their cytotoxicity to their abihty to Interact with, and damage, DNA. Many agents form bulky adducts that block RNA and DNA polymerases, inhibiting transcription, and DNA rephcation.

Immunofluorescent staining techniques can be used to probe the DNA structure of chromosomes. The primary requirement IS a sequence- or structure-specific antrbody. Most DNA is not immunogeinc, although DNA-binding antibodies are produced spontaneously in the autoimmune drsea ...

DNA-binding antibodies are produced spontaneously in the autoimmune disease systemic lupus erythematosus (1). The origin of these antibodies is obscure, but they are involved in the pathogenesls of the disease through the formation of immune complexes that indirectly cause tissue d ...

Complexation between a ligand molecule and a nucleic acid leads to optical changes that can be used to monitor the binding process. As these host—drug interactions frequently involve a reversible mechanism, a determination of the equilibrium binding constant can provide insight into t ...

If the temperature of a solution containing a helical nucleic acid is raised sufficiently, strand separation, or “melting,” occurs (1–3) (Fig. 1) (see Note 1) The temperature that marks the midpoint of the melting process is called the melting temperature (or T m) (Fig. 2). At the T m, half of the nucleic acid exi ...

Among the methods avallable for the analysis of structures of drug—DNA complexes in solution, the electric dichrolsin is particularly simple, does not require much material, and provtdes information, which cannot be obtained as conveniently by other methods The principle of the elect ...