Incorporation of radiolabeled

Measured end points are photosynthesis as the incorporation of radiolabelled H C03 ( C) and bacterial activity as the incorporation of radiolabelled thymidine (thym) fluorometric measurements basal fluorescence (Fo) and photon yield (Y) chlorophyll-a concentration (chl-a) species composition (spp) and the biovolume of algae obtained after algal counting (biovolume)... 

The immobilized immunoprecipitates are washed twice with lysis buffer containing 0.5 MNaCl and twice with buffer A. The beads are resuspended in 20 /il of kinase buffer also containing the appropriate concentration of the specific peptide. Reactions should also be set up without peptide as a negative control for nonspecific or self-incorporation of radiolabel. To start the reactions, 5 /il of ATP is added (final concentration 0.1 mM unlabeled ATP, 1 /iCi [7 -32P]ATP (per assay) in kinase buffer). The assays are allowed to proceed for 15 to 30 min at 30° with constant shaking at 900 rpm, and stopped by spotting 20 /il of the sample (slurry) onto a square (1.5 X 1.5 cm) of phosphocellulose (P81) paper. The P81 papers are immediately immersed in 500 ml of 1% (v/v) orthophosphoric acid, and then washed 3 times with the same solution (to remove the excess ATP). The washes therefore contain almost all of the radiolabel and must be handled carefully and disposed of appropriately. The papers are briefly rinsed in ethanol and air-dried. The incorporation of 32P-label is measured by Cerenkov counting. 

Dobbins DC, Pfaender FK. 1988. Methodology for assessing respiration andcellular incorporation of radiolabeled substrates by soil microbialcommunities. Microb Ecol 15 257-273. 

In relation to their biosynthesis, it was proposed that A-B units of Et s are most likely formed by condensation of two Dopa-derived building blocks and that the tetrahydroisoquinoline ring in unit B is closed by condensation (Pictet-Spengler) with a serine(or glycine)-derived aldehyde, as in the case of the related saframycins. S-Adenosylmethinonine is the likely source of the methyl groups. A plausible route for the formation of unit C was proposed later [75]. This was partially demonstrated by incorporation of radiolabeled tyrosine and cysteine by Kerr and Miranda [80] and by incorporation of labeled methionine, glycine and tryptophan by Morales and Rinehart [81]. 

Bjostad and Roelofs (1983) were the first to determine correctly how the major pheromone component for a particular moth was biosynthesized. This was done by looking for possible fatty acid intermediates and by monitoring the incorporation of radiolabeled precursors into pheromone components. They showed that glands of the cabbage looper, Trichoplusia ni, utilize acetate to produce the common fatty acids octadecanoate and hexadecanoate which undergo All desaturation to produce Zll-18 acid and Zll-16 acid. But the main pheromone component was Z7-12 OAc, which presumably was made from Z7-12 acid. To demonstrate how the fatty acid precursor Z7-12 acid was produced, [3H-16]-Z1 l-16 acid was applied to glands and monitored for incorporation. It was incorporated into both Z7-... 

Jeffrey pine beetle, Dendroctonus jeffreyi Hopkins, which had been previously treated with juvenile hormone III (JH III, 2.2 pg/beetle in acetone) and then placed in an aeration tube for 25 to 30 h. Ips paraconfusus and I. pini were each injected with 0.2 pCi of sodium [1-14C]acetate prior to placement in cut pine logs and volatile collection, while D. jeffreyi were each injected with 3.8 (male) and 3.7 (female) pCi of sodium [1-14C]acetate 6.4 (male) and 10.7 (female) h after JH application. (G) The role of the mevalonate pathway in frontalin biosynthesis is supported by the incorporation of radiolabel from [2-14C]mevalonolactone into frontalin by male D. jeffreyi (2.2 pg JH 11 l/beetle in acetone, 10 h incubation and volatile collection, 1.1 pCi of [2 14C] mevalonolactone injected, 20 h volatile collection). Figures adapted from Seybold et al. (1995b) and Barkawi (2002). 

The first argument was supplied by Post and Vincent (4o) in their study of the incorporation of radiolabeled glucose in tissue fractions of normal... 

The upper-left panel of figure 6.3 shows how variations in the concentrations of KC1 or K-acetate affect the incorporation of radiolabeled lysine into newly synthesized proteins in mouse L-cells (Weber et al., 1977). Both potassium salts stimulate protein synthesis up to a certain salt concentration, with the acetate salt having a higher optimal concentration and higher final degree of stimulation than KC1. Above the opti-... 

Nucleic acid probes are commonly labeled by enzymatic incorporation of radiolabeled nucleotides or enzymatic addition of radiolabeled phosphate groups to the nucleic acid chain. Proteins, in particular immunoglobulins, are labeled commonly by direct... 

P. vitcola and many other oomycetes are wide-spread in field populations, and fluctuate in frequency over years and within seasons but are in a dynamic equilibrium with sensitive isolates in many areas [55]. Inheritance of resistance was demonstrated to be monogenic with the participation of minor genes [56]. Biochemical studies (incorporation of radiolabeled uridine) suggested that a change of RNA polymerase I is responsible for resistance [54], However, only recently sequence data for polymerase subunits became available in P. infestans [57] but molecular detection tools have not yet been developed. 

Effect of Sarin on DNA and Protein Content Kassa et al (2000) showed that not only symptomatic level 3 but also asymptomatic levels 1 and 2 of sarin were able to significantly decrease the incorporation of radiolabeled thymidine without changing total concentrations of DNA or... 

In contrast to the rapid changes upon receptor activation in intracellular concentrations of lower inositol polyphosphates (e.g., Ins(l,4,5)P3), InsPe concentrations seem immutable. This finding, along with observations of the sluggish incorporation of radiolabeled inositol into InsPe, prompted interpretations of slow turnover rates for InsPe in vivo. However, the identification of InsPe as a precursor to even more phosphorylated inositol polyphosphates overturned these misconceptions and revealed the true dynamic nature of cellular InsPe. 

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