CDNA data

Material collection —> mRNA microarray (GeneChip or CDNA) — Data mining tools <-> Drug Discovery... 

In principle, this is a straightforward procedure for sequencing proteins. Like any other methodology, this procedure has its pitfalls and care should be exercised, especially in the analysis of novel proteins and peptides for which other information (e.g., cDNA data) is not available. 

Machado, O.L.T., Silva Jr., J.G., 1992. An allergenic 2S storage protein from Ricinus communis seeds which is a part of the 2S albumin precursor predicted by cDNA data. Braz. J. Med. Biol. Res. 25,567-582. 

FIGURE 3.11 Constitutive activity in melanophores expressing hCTR2 receptor, (a) Basal melanophore activity, (b) Effect of transfection with human cDNA for human calcitonin receptors (16 j-ig/ml). (c) Concentration response curve for cDNA for human calcitonin receptors (abscissae as log scale) and constitutive activity. Data redrawn from [27]. 

FIGURE 3.12 Dependence of constitutive receptor activity as ordinates (expressed as a percent of the maximal response to a full agonist for each receptor) versus magnitude of receptor expression (expressed as the amount of human cDNA used for transient transfection, logarithmic scale) in Xenopus laevis melanophores. Data shown for human chemokine CCR5 receptors (open circles), chemokine CXCR receptors (filled triangles), neuropeptide Y type 1 receptors (filled diamonds), neuropeptide Y type 2 receptors (open squares), and neuropeptide Y type 4 receptors (open inverted triangles). Data recalculated and redrawn from [27],... 

The cDNA encoding the luciferase of Renilla reniformis has been obtained and expressed in Escherichia coli (Lorenz et al., 1991). The cDNA contained an open reading frame encoding a 314-amino acid sequence. The recombinant Renilla luciferase obtained had a molecular weight of 34,000, and showed an emission maximum at 480 nm in the luminescence reaction of coelenterazine, in good agreement with the data of natural Renilla luciferase. 

Sequences of the genes/cDNAs can be retrieved from databases on the Internet at various web sites. For example, GeneBank (at the National Center for Biotechnology Information, NCBI) is at http //www.ncbi.nlm.nih.gov/ Web/Search/index.html. The EMBL Nucleotide Sequence database (through the European Bioinformatic Institute, EBI) can be found at http //www.ebi.ac.uk/queries/queries.html, whilst that of the DNA Data Bank of Japan is at http //www.ddbj.nig.ac.jp/. 

The PE2 isoform has been purified and fully sequenced (Markovic and Joumval 1986). Using this sequence data Ray et al (1988) succeeded in isolating a clone fi om a tomato fruit cDNA library. The predicted amino acid sequence fi om this clone had high homology to the actual amino acid sequence determined for PE2 but was not identical. Subsequent screening of the tomato... 

Other recent and very compelling evidence that the P subunit may also be necessary for full activity [83] comes from the study of a high threshold but DHP-insen-sitive brain channel that has an a 1-like channel core. The cDNA for this brain channel was recently isolated and the mRNA was expressed in Xenopus oocytes. The current corresponding to this channel was increased when the mRNA for either the skeletal muscle 0.2 or P subunits were co-injected. However, when the brain ai-like mRNA was co-expressed with the combination of the skeletal muscle 2 and p mRNAs, the current was dramatically increased from 31 nA for the brain ai alone to 6 500nA for the combination [83]. These striking results are the best evidence so far obtained that the P subunit has a functional role in channel activity. Although these data were obtained with a DHP-insensitive channel, they pro-... 

Evidence for de novo synthesis of pheromone components was obtained by showing that labeled acetate and mevalonate were incorporated into ipsdienol by male Ips pini [103,104]. Similarly, labeled acetate and other labeled intermediates were shown to be incorporated into frontalin in a number of Dendroctonus species [105]. Possible precursors to frontalin include 6-methyl-6-hep-ten-2-one, which was incorporated into frontalin by D. ruffipennis [106]. The precursor 6-methyl-6-hepten-2-one also was shown to be converted to bre-vicomin in the bark beetle, Dendroctonus ponderosae [107]. In addition, the expression patterns of HMG-CoA reductase and HMG-CoA synthase are tightly correlated with frontalin production in Dendroctonus jeffreyi [108, 109]. A geranyl diphosphate synthase cDNA from I. pini was also isolated, functionally expressed, and modeled [110]. These data indicate that the de novo isoprenoid biosynthetic pathway is present in bark beetles. A variety of other monoterpene alcohols such as myrcenol, pityol, and sulcitol are probably synthesized through similar pathways [111]... 

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