Polymerase chain reaction synthases

ACC 71 synthase, i. e. (S)-adenosylmethionine methylthioadenosine lyase (EC 4.4.1.14), has been purified from several plant tissues [116]. Recently, ACC synthase cDNA clones have been isolated and sequenced from wounded fruit tissues of tomato, winter squash, zucchini, ripening apple and tomato fruit. Using the polymerase chain reaction (PCR), four different ACC synthase gene fragments were obtained by amplification of cDNA derived from mRNA of tomato... 

Bracher, D. and Kutchan, T. M. 1992. Polymerase chain reaction comparison of the gene for strictoside synthase from ten Rauvolfia species. Plant Cell Reports, 11 179-182. 

The phbA, phbB, and phbC genes from Alcaligenes eutrophus (Ralstonia eutrophus) encoding the biosynthetic enzymes (3-ketothiolase, acetoacetyl-CoA reductase (NADPH-dependent), and PHB synthase, respectively, have been cloned into E. coli (Scheme 19.42).339-342 The use of in vitro evolution using error-prone polymerase chain reaction has led to enhanced accumulation of PHA in a resultant recombinant strain.343 Additional studies to enhance the biosynthesis of PHB through the use of metabolic engineering have been discussed.344... 

Solaiman, D. K. Y., Ashby, R. D., and Foglia, T. A. 2000. Rapid and specific identification of medium-chain-length polyhydroxyalkanoate synthase gene by polymerase chain reaction. Appl. Microbiol. Biotechnol., 53, 690-694. 

Solaiman, D. K. Y. 2002. Polymerase-chain-reaction-based detection of individual polyhydroxyalkanoate synthase phaCl and phaC2 genes. Biotechnol. Lett., 24, 245-250. 

Abbreviations used EMSA, electrophoretic mobility shift analysis IMPase 1, inositol monophosphatase 1 inositol synthase, A/vo-inositol 3-phosphate synthase NCBI, National Center for Biotechnology Information PCR, polymerase chain reaction Rb, retinoblastoma protein tss, transcriptional start site IP3, Inositol trisphosphate DAG, diacyl glycerol FISH, fluorescent in situ hybridization. 

Terada, Y., Tomita, K., Nonoguchi, H., and Marumo, F. (1992). Polymerase chain reaction localization of constitutive nitric oxide synthase and soluble guanylate cyclase messenger RNAs in microdissected rat nephron segments. J. Clin. Invest. 90, 659-665. 

Another important application arising from the work with trichodiene synthase has been the development of Tri5 specific polymerase chain reaction-based assays for the detection of potential trichothecene-producing Fusarium species in pure culture and in contaminated grain (269,275,276). 

Inducible nitric oxide synthase (iNOS EC 1.14.13.39) mRNA in bronchoalvelar lavage cells (using reverse transcriptase-polymerase chain reaction) and the magnitude of NO-dependent chemiluminescence from alveolar macrophages correlated with the pathology of silica-exposed coal miners chest radiographs (Castranova et al. 1998). 

Haematology DNA damage in peripheral blood mononuclear cells and total hydrophilic antioxidant capacity in the plasma from 19 TB patients and healthy tuberculin test-positive controls were evaluated by single-cell gel electrophoresis and inducible isoform Nitric Oxide Synthase (iNOS) expression measured by quantitative polymerase chain reaction (qPCR). Compared to controls, pulmonary TB patients under treatment presented with increased DNA damage and increased antioxidant capacity, which diminished during treatment. TB patients showed lower iNOS expression, but expression tended to increase during treatment [36 ]. 

The mechanism for bacterial synthesis of PHA is not the simple dehydration reaction between alcohol and carboxyl groups. It is more complicated and involves the coenzyme A thioester derivative of the hydroxyalkanoic acid monomer (produced from the organic feedstock available to the bacteria) [Kamachi et al., 2001]. Growth involves an acyl transfer reaction catalyzed by the enzyme PHA synthase (also called a polymerase) [Blei and Odian, 2000]. The reaction is not a step polymerization but is a chain polymerization with the characteristics of a living polymerization [Su et al., 2000] (Secs. 3-15, 5-2g, Chap. 7). 

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