Restriction enzyme analysis applications

Most often proteins are the bacterial biopolymers studied using MALDI MS either from fractions or whole cells. They are not the only isolated cellular biopolymers studied by MALDI, nor the first. Very soon after the introduction of MALDI there were a few reports of the analysis of bacterial RNA or DNA from bacterial fractions. One of the first applications of MALDI to bacteria fractions involved analysis of RNA isolated from E. coli,4 Other studies included analysis of PCR-amplified DNA,5 6 DNA related to repair mechanisms7 and posttranscriptional modification of bacterial RNA.8 While most MALDI studies involve the use of UV lasers, IR MALDI has been reported for the analysis of double stranded DNA from restriction enzyme digested DNA plasmids, also isolated from E. coli.9... 

Hsieh, Y.W. 2003. Studies on application of restriction enzyme and analysis in partial cytochrome b gene for species identification of puffer fish and related products. Ph.D. Thesis, National Taiwan Ocean University, Keelung. p. 191. 

The use of restriction enzymes to cleave DNA at specific sequences was mentioned earlier in this chapter in the context of DNA sequence analysis. These enzymes are also important in the field of recombinant DNA technology. We will illustrate this application by describing a method for the production of human insulin. 

Wada, Y. (1998) Separate analysis of complementary strands of restriction enzyme-digested DNA. An application of restriction fragment mass mapping by matrix-assisted laser desorption/ ionization mass spectrometry. J. Mass Spectrom., 33 (2), 187 192. 

This restricts the use of immobilized whole cells in analysis of single substances to three main applications a) determination of a substance in a pure product stream when production of by-products does not matter as long as that part of the total metabolism involved is constant, thus giving a reproducible signal b) use of organisms with increased amounts of a specific enzyme or c) mutants that have completely lost one enzyme function. 

In the world market, more than 90% of commercial biosensors are enzymatic, in particular, those that measure glucose, used by diabetics [79]. This kind of biosensors is very useful because if immobilized enzymes are sensitive to certain pollutants, these analytes can be easily measured. For example, biosensors based on the inhibition of acetylcholinesterase detect phosphorus insecticides and other inhibitors [80]. A comprehensive analysis showed that the developed enzymatic biosensors demonstrated reproducible, stable, and fast responses to the substrates to be measured. Unfortunately, the application of these biosensors can be restricted because of the dramatic decrease in the sensor response at increasing buffer capacity and ionic strength, pH-dependence of the enzyme kinetics, and cosubstrate limitation of the measured enzymatic reaction rate (the glucose sensor) [79]. Recently, Soldatkin et al. reported a complete review of some biopolymers used in enzymatic... 

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