化学生物学实验技术

相关专题 细胞培养常用试剂指南胎牛血清产品选择指南 【目的要求】1. 掌握高铁-硫酸显色法测定血清总胆固醇的原理和方法。2. 了解血清胆固醇的正常值及临床意义。【实验原理】胆固醇（cholesterol）是环戊烷多氢菲的衍生物，它不仅参与血浆蛋白的组成，而且也是细胞的必要结构成分，在神经组织和肾上腺中含量特别丰富，在肝、肾和表皮组织含量也很多。胆固醇是生物膜和神经髓鞘的重要组成部分，是维持生物膜的正常透过能力不可缺少的。胆固醇还可 ...

The interaction of ligands with G protein-coupled receptors (GPCRs) has been traditionally studied using radiolabelled variants of receptor ligands. More recently, increased knowledge about the way in which GPCRs exist in a highly organised membrane environment has led to an intere ...

Gene therapy represents a set of approaches to the treatment of diseases based on the transfer of genetic material (DNA) into an individual (or animal) and is defined as the use of nucleic acid transfer, either RNA or DNA, to treat or prevent a disease 1–3). The process involves a group of technologies that en ...

Inborn errors of metabolism are inherited conditions where there is a defect or block in a metabolic pathway leading to the accumulation of metabolite(s) proximal to the defect and a metabolic deficit distal to this (1). The defect usually results from a specific enzyme deficiency; however, def ...

The completion of the Human Genome Project (HGP) was announced on 24 April 2003 (1). It was a landmark event in the history of biology and represents one of the most remarkable achievements in the history of science and signals the beginning of a new era in biomedical research. This chapter will describe br ...

The sequencing of the human genome has opened the door for proteomics by providing a sequence-based framework to mine the human proteome. Although the field of proteomics was initially dominated by two-dimensional gels and mass spectrometry, the current emphasis is on developing proteo ...

For a considerable fraction of patients who acquire a bacterial infection during their stay in the hospital (i.e., a nosocomial infection), the infection was acquired through clonal dissemination of the bacteria from another patient or from a hospital source. Alternately, the source of the ...

Capillary electrophoresis (CE) is an exciting technique that has many clinical applications. These include serum and urine protein electrophoresis, the diagnosis of hemoglobinopathies, the quantification of drugs such as iohexol and phenobarbital, and the analysis of DNA.

Chromosomal translocations were the first target for the specific detection of residual tumor cells in bone marrow and peripheral blood. Some types of leukemia are regularly or generally associated with translocations. In chronic myelogenous leukemia (CML) and a proportion of pati ...

Advances in molecular genetics and biotechnology are changing the practice of medicine. Completion of the human genome sequence and development of high-throughput semiautomated DNA analysis techniques have resulted in an exponential increase in the rate of discovery of new disea ...

Nucleic acid sequence-based amplification (NASBA; bioM�rieux, Boxtel, The Netherlands) is a commercially available amplification procedure that uses RNA as the target. It makes use of the simultaneous enzymatic activities of avian myeloblastosis virus reverse transcripta ...

It has been estimated that the human genome contains 25,000–30,000 genes that are distributed on the 24 chromosomes. Therefore, the identification and localization of any particular gene responsible for a given trait or disease has always been a difficult task.

Nucleic acid amplification technologies have greatly facilitated medical diagnostics for genetic and infectious diseases through the exquisite sensitivity and specificity associated with these methods. Polymerase chain reaction (PCR) (see Chapter 6) ushered in these te ...

Allele-specific polymerase chain reaction (PCR) was first described in 1989, with variations arising over the next few years such as allele-specific oligonucleotide PCR, mutant-allele-specific amplification (MASA), PCR amplification of specific alleles (PASA), and the ampl ...

Conformation-sensitive gel electrophoresis (CSGE) was initially developed by Ganguly et al. in 1993 as a screening method to minimize the amount of nucleotide sequencing required when investigating large multiexon genes for mutations (1). It is based on the ability to distinguish bet ...

Screening for mutations of the thousands of the sequence products provided by human genome analysis has proven to be a daunting task. The gold standard for identifying sequence alterations is direct sequencing. However, this method is labor- intensive and the least costeffective. Since t ...

Capillary electrophoresis (CE) separations are carried out inside a capillary tube, which usually has a diameter of 50 μm to facilitate temperature control. The length of the capillary differs in different applications, but it is typically in the region of 20–50 cm. The capillaries most widely ...

The identification of a large number of disease genes in recent years has led to a considerable improvement in clinical diagnostic procedures, therapeutic interventions, and prognostic projections and provided carrier or presymptomatic testing to family members of affected ind ...

Restriction enzymes (or restriction endonucleases) are bacterial enzymes capable of cleaving double-stranded DNA. Even though the enzymes are bacterial in origin, because of the universal nature of DNA they can digest DNA from any species, including humans. Importantly, restrict ...

There have been many developments over the past three decades that have led to the efficient manipulation and analysis of nucleic acid and proteins. Many of these have resulted from the isolation and characterization of numerous DNA-manipulating enzymes, such as DNA polymerase, DNA ligas ...