Biochemistry. the molecular basis of life
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Biochemistry: The Molecular Basis of Life, Fourth Edition, is the ideal text for students who do not specialize in biochemistry but require a strong grasp of the essential biochemical principles of the life and physical sciences for their future careers. Specifications Publisher Oxford Univ Pr. Customer Reviews.
Biochemistry : the molecular basis of life
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Aimed at students who have a previous knowledge of organic chemistry, the text focuses on essential biochemicalprinciples that underpin the modern life sciences, and offers the most balanced coverage of chemistry and biology of any text on the market. Biochemistry: The Molecular Basis of Life provides a complete view of the living state by explaining the functional and structural properties of biomoleculesin the context of their biochemical reactions and impact on living organisms.
DNA samples before or after restriction enzyme restriction endonuclease digestion are separated by gel electrophoresis and then transferred to a membrane by blotting via capillary action. The membrane is then exposed to a labeled DNA probe that has a complement base sequence to the sequence on the DNA of interest.
These blots are still used for some applications, however, such as measuring transgene copy number in transgenic mice or in the engineering of gene knockout embryonic stem cell lines. The northern blot is used to study the expression patterns of a specific type of RNA molecule as relative comparison among a set of different samples of RNA. It is essentially a combination of denaturing RNA gel electrophoresis , and a blot. In this process RNA is separated based on size and is then transferred to a membrane that is then probed with a labeled complement of a sequence of interest.
The results may be visualized through a variety of ways depending on the label used; however, most result in the revelation of bands representing the sizes of the RNA detected in sample. The intensity of these bands is related to the amount of the target RNA in the samples analyzed.
The procedure is commonly used to study when and how much gene expression is occurring by measuring how much of that RNA is present in different samples. It is one of the most basic tools for determining at what time, and under what conditions, certain genes are expressed in living tissues. In western blotting , proteins are first separated by size, in a thin gel sandwiched between two glass plates in a technique known as SDS-PAGE.
The proteins in the gel are then transferred to a polyvinylidene fluoride PVDF , nitrocellulose, nylon, or other support membrane. This membrane can then be probed with solutions of antibodies. Antibodies that specifically bind to the protein of interest can then be visualized by a variety of techniques, including colored products, chemiluminescence , or autoradiography.
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- Trudy McKee and James McKee, Biochemistry: The Molecular Basis of Life 7e.
Often, the antibodies are labeled with enzymes. When a chemiluminescent substrate is exposed to the enzyme it allows detection. Using western blotting techniques allows not only detection but also quantitative analysis. Analogous methods to western blotting can be used to directly stain specific proteins in live cells or tissue sections. The eastern blotting technique is used to detect post-translational modification of proteins. Proteins blotted on to the PVDF or nitrocellulose membrane are probed for modifications using specific substrates. A DNA microarray is a collection of spots attached to a solid support such as a microscope slide where each spot contains one or more single-stranded DNA oligonucleotide fragments.
Arrays make it possible to put down large quantities of very small micrometre diameter spots on a single slide. A variation of this technique allows the gene expression of an organism at a particular stage in development to be qualified expression profiling. This cDNA is then hybridized to the fragments on the array and visualization of the hybridization can be done. Since multiple arrays can be made with exactly the same position of fragments they are particularly useful for comparing the gene expression of two different tissues, such as a healthy and cancerous tissue.
Also, one can measure what genes are expressed and how that expression changes with time or with other factors. There can be anywhere from spots to more than 10, on a given array. Arrays can also be made with molecules other than DNA. Allele-specific oligonucleotide ASO is a technique that allows detection of single base mutations without the need for PCR or gel electrophoresis. Short 20—25 nucleotides in length , labeled probes are exposed to the non-fragmented target DNA, hybridization occurs with high specificity due to the short length of the probes and even a single base change will hinder hybridization.
The target DNA is then washed and the labeled probes that didn't hybridize are removed. The target DNA is then analyzed for the presence of the probe via radioactivity or fluorescence. In this experiment, as in most molecular biology techniques, a control must be used to ensure successful experimentation. In molecular biology, procedures and technologies are continually being developed and older technologies abandoned. For example, before the advent of DNA gel electrophoresis agarose or polyacrylamide , the size of DNA molecules was typically determined by rate sedimentation in sucrose gradients , a slow and labor-intensive technique requiring expensive instrumentation; prior to sucrose gradients, viscometry was used.
Aside from their historical interest, it is often worth knowing about older technology, as it is occasionally useful to solve another new problem for which the newer technique is inappropriate. While molecular biology was established in the s, the term was coined by Warren Weaver in Weaver was the director of Natural Sciences for the Rockefeller Foundation at the time and believed that biology was about to undergo a period of significant change given recent advances in fields such as X-ray crystallography.
Clinical research and medical therapies arising from molecular biology are partly covered under gene therapy. The use of molecular biology or molecular cell biology approaches in medicine is now called molecular medicine.
Biochemistry: The Molecular Basis of Life - Trudy McKee, James Robert McKee - Google книги
Molecular biology also plays important role in understanding formations, actions, and regulations of various parts of cells which can be used to efficiently target new drugs , diagnose disease, and understand the physiology of the cell. From Wikipedia, the free encyclopedia. Branch of biology dealing with biological activity's molecular basis.
For the scientific journal, see Biochemical Genetics. Index Outline.
For more extensive list on protein methods, see protein methods. For more extensive list on nucleic acid methods, see nucleic acid methods. Main article: Molecular cloning. Main article: Polymerase chain reaction. Main article: Gel electrophoresis. Main article: Southern blot. Main article: Northern blot. Main article: Western blot.