生物化学技术

The aim of this chapter is to give a general overview of the technique of in situ hybridization By way of introducing the technique, the history and background of in situ hybridization will first be discussed, followed by the principles and basic steps involved and lastly, the various applications ...

Mass spectrometry (MS) (1) is one of the most important physical methods in analytical chemistry today A particular advantage of mass spectrometry, compared with other molecular spectroscopies, is its high sensitivity, so that it provides one of the few methods that is entirely suitable for ...

The characterization of gene expression patterns within cells and tissues is a very desirable and highly informative way of providing insights into cell development, maintenance, and cell-cell interactions. Several widely used methods, such as Northern or dot blot, in situ hybridiza ...

By defimtion, nnmunocytochemistry IS a biomolecular technique that mvolves the use of a specific antibody-antigen reaction to identify cellular constituents in situ The techmque requires that the antibody be tagged with a label to facilitate its visualization Immunocytochemi ...

An epitope can be simply defined as that part of an antigen involved in its recogmtion by an antibody In the case of protein antigens, an epitope would consist of a group of mdlvidual ammo-acid side-chains close together on the protein surface Epitope mapping, then, becomes the process of locating the e ...

Foreign proteins can be displayed on the surface of filamentous bacteriophage by fusing them to the phage minor coat protein (Fig. 1) Many proteins have been expressed in this way, including human growth factor (1), alkaline phosphatase (2), ricin (3), pancreatic trypsin inhibitor (4), and a DNA-b ...

It is approaching half a century since Edman developed a chemistry for the determination of the N-terminus of a protein or peptide (1), and thts chemistry is still being used today. in the intervening decades various ancillary techniques have come (and some have gone) that have allowed the field to de ...

The analysis of RNA molecules by S1 nuclease digestion constitutes an important tool for molecular studies of cellular processes, not only because of the historical significance of its contribution to messenger ribonucleic acids (mRNA) structure analyses, but also for its simplici ...

Reversed-phase HPLC (RP-HPLC) is now well establrshed as a technique for rsolanon, analysts, and structural elucrdatron of peptrdes and proteins (1,2) Its use m protem rsolatron and purificatron may have reached a peak owing to recent developments m high efficiency ron-exchange and hydro ...

Protein blotting was initially introduced in the late 1970s to identify protein antigens that bound to spectfic antibodies (1,2) In this procedure, a complex protein fraction was separated by electrophoresis and the proteins transferred and bound to a membrane, which was then probed with a r ...

Electrophoresls wlthm silica caplllarles is a relatively new technique, with the first pubhcatlons appearing in the 1970s followed by a rapid expansion and establishment of the technique in the 1980s Commercial machines for the apphcatlon of capillary electrophoresis appeared in ...

Radioacttve labeled pepttdes and proteins are used extensively in many areas of biochemtstry, pharmacology, and medicme For example, they are frequently employed as tracer molecules in quantitative determmattons, such as measurement of hormone and hormone-receptor concent ...

Of the vast range of techniques in use in modern molecular biology, DNA protiling is one of the more difficult to describe, primarily because the term “DNA Protiling” itself means very different things to different people. It is, in fact, a widely applied term that describes a number of technically div ...

The ability to create defined mutations and to generate precise fusions of protein domams by modifications at the DNA level is now an accepted and routine technique in molecular biology This ability has developed over the past 10–l5 years and rests on a foundation of experiments on DNA.DNA hybrid ...

The process of transcriptron in which a DNA template is used to derive RNA copies is a fundamental process in prokaryotic and eukaryotic cells The recaprtulatron of these processes in cell-free systems (m vitro transcription) has led to a number of powerful techniques for studying gene struct ...

Since their mtroductlon in 1987, yeast artificial chromosomes (commonly referred to as YACs) have had a tremendous impact on the field of molecular genetics The ability of YACs to carry and propagate DNA fragments of up to -2 mllhon base pairs in size has been instrumental in the construction of the fi ...

The genome maps of a small number of prokaryotes have been completed. In recent years, efforts have been made to map the entire mouse and human genomes; indeed the human-genome project-a global effort to clone, sequence, and map the human genome-represents the most ambitious and challenging task of ...

Knowledge of eukaryotic-gene structure has grown exponentially over the past two decades, mainly owing to the mtroduction of new molecular techniques These have enabled the manipulation and analysis of the DNA molecule and led to the identificatlon of mutations causing single gene dis ...

Plant transformation involves the insertion of “foreign” genetrc material into plant cells and the regeneration of transgenic plants Over the past 15 yr or so, a variety of methods for plant genetic engineering have been described, these fall into two broad categories. Agrobactenum-bas ...

The transfer or “transfectron” of genetic material into tissue-culture cell lines and the subsequent analysis of its RNA and protein products, has become a key and central technology in contemporary cell biology, molecular biology, and protein chemtstry. Transfectron of genes provid ...