细胞功能测定

The proteoglycans are a group of macromolecules characterized by the covalent attachment of one or more sulfated glycosaminoglycan (GAG) chains to a protein core, and are an important constituent of the extracellular matrix. Cartilage contains a number of proteoglycans, the most abun ...

Degradation of fibrillar collagens by matrix metalloproteinases (MMPs) is thought to be a major catabolic pathway in various connective tissues. However, the complex structure of collagen molecules and their degradation products makes specific assay of these events very challe ...

Matrix metalloproteinases are believed to play a major role in cellular invasion and tumor metastasis. Therefore, to understand the molecular mechanisms underlying invasion, a variety of in vitro assays have been developed. Most assays assess the interaction of tumor cells with the sur ...

Catabolism of extracellular matrix (ECM) components has been ascribed to a family of Zn2� metalloenzymes. These matrix metalloproteinases (MMPs; also termed matrixins) are believed to be important in connective tissue remodeling during development and wound healing. MMPs have al ...

Tissue inhibitors of metalloproteinases (TIMPs) are well known as inhibitors of matrix metalloproteinases (MMPs) such as interstitial collagenase, gelatinase A and B, and stromelysin 1, and have been suggested to play an important role in the regulation of MMPs. Although MMP inhibition ...

The kinetic analysis of the inhibition of matrix metalloproteinases by tissue inhibitor of metalloproteinases (TIMP) yields valuable information on the mechanism and specificity of the TIMPs. When combined with the use of genetic engineering or chemical methods of modification to ...

The emergence of cycle sequencing (1) as a powerful alternative to conventional isothermal methods has facilitated the manipulations involved in sequencing protocols in general, and is of value for the sequence analysis of PCR products, in particular. Using radioactive labeling tec ...

At least 19 proteins have now been defined as collagens (1,2), but many of those recently discovered are present in tissues in such small amounts that their isolation for characterization at the protein level has so far been impossible. Some of the fibril-forming collagens are now in medical use, in ap ...

Techniques for direct sequencing of PCR products are of central importance to contemporary research in molecular biology and genetics. The rapidly growing number of cloned human disease genes increasingly allows sequencing of PCR amplicons for diagnostic purposes. Nonradioac ...

The conventional methods of sequencing, such as Maxam and Gilbert (1) (involving chemical cleavage of labeled DNA fragments) and Sanger et al. (2), of dideoxy sequencing (using enzymatic extension of oligonucleotide primers) remain the most reliable techniques, but when compared to more ...

The polymerase chain reaction (PCR) allows the rapid detection of infectious viruses or other microorganisms as well as the cause of genetic defects. By performing sequence analysis afterward, important additional information on the PCR products is obtained. Often direct sequenci ...

The polymerase chain reaction (PCR) has become widely established as a powerful core molecular biology technique because of its ability of produce large amounts of specific target DNA from limited template sources (1). Numerous applications based on the PCR have also been developed, incl ...

Among the many techniques of cloning new genes, one approach involves degenerate primers (1–7). The approach usually requires three steps: 1. Using degenerate primers to amplify part of the gene of interest by PCR: The degenerate primers’ sequence

The inherent problems of sensitivity and specificity that one encounters when trying to determine a particular nucleotide sequence directly in its genomic context can be overcome by selective amplification of the region of interest. This amplification of the target DNA is usually ach ...

The advance of Taq-based polymerase chain reaction (PCR) technology (1–3) has had a tremendously positive impact on biomedical research. The combination of PCR and sequencing further revolutionized biological research (3, 4). Sequence amplification technology has provided a spe ...

The acceptance of polymerase chain reaction (PCR) as a routine method in molecular biology has created a need for simple and robust approaches to DNA sequencing of the amplification products. In addition to its ease, direct sequencing of PCR products has the advantage that nucleotide misinc ...

The polymerase chain reaction (PCR) is well known for being a rapid and versatile method for the amplification of defined target DNA sequences. This technique can be applied to a variety of research areas, such as the identification and typing of single nucleotide substitutions of DNA sequence ...

The use of the polymerase chain reaction (PCR) allows isolation and examination of genomic DNA sequences using specific oligonucleotide primers to amplify a target region with Taq DNA polymerase (1,2). The PCR products can be screened for mutations using molecular techniques, such as all ...

The polymerase chain reaction (PCR) facilitates the rapid in vitro amplification of target DNA segments. Numerous applications have been developed to exploit the vast potential of PCR, and many of these require sequence analysis of the DNA product. This chapter describes the dideoxy chai ...

Although numerous methods are now available for direct sequencing of PCR products, cloning of amplified DNA for sequencing in M13 vectors remains an attractive approach because of the high quality of sequence information generated from single-stranded bacteriophage DNA templat ...