Eukaryotes analysis

Volume 449. R NA Turnover in Eukaryotes Analysis of Specialized and Quality Control RNA Decay Pathways... 

The existence of two separate but interacting photosystems in photosynthetic eukaryotes was demonstrated through analysis of the photochemical action spectrum of photosynthesis, in which the oxygen-evolving capacity as a function of light wavelength was determined (Figure 22.10). 

Structural analysis of the rhinovirus and the hepatitis A virus 3C proteases (Allaire et al. 1994 Matthews et al. 1994) confirmed earlier predictions that the picomavirus 3C proteases are similar to chymotrypsin-Uke serine proteases in their fold. An important difference is that the serine nucleophile of serine proteases is replaced with a cysteine however, the 3C protease is stracturally distinct from the eukaryotic cysteine protease class of enzymes. 

Gentz, R., Hayes, A., Grau, N. et al. (1992) Analysis of soluble human and mouse interferon-gamma receptors expressed in eukaryotic cells. European Journal of Biochemistry, 210 (2), 545-554. 

Fig. 5.1. The three domains of life in a radial time sequence based on genetic (RNA) analysis. The distinction between anaerobes and aerobes is not made here and the branching to eukaryotes is left unclear. All the domains advanced with time but in very different ways (see Woese, C. (1998) in Further Reading, and Sogin, M. (1993), Science, 260, 340). Note the general gene transfer in prokaryotes and to eukaryotes (see Chapter 7).

Before we describe the chemistry of the compartments involved, note that like prokaryotes, a number of oxidative enzymes are found in the cytoplasm but they do not release damaging chemicals (see Section 6.10). We also observed that such kinds of kinetic compartments are not enclosed by physical limitations such as membranes. We have also mentioned that increased size itself makes for kinetic compartments if diffusion is restricted. In this section, we see many additional advantages of eukaryotes from those given in Section 7.4. How deceptive it can be to use just the DNA, the all-embracing proteome, metabolome or metallome in discussing evolution without the recognition of the thermodynamic importance of compartments and their concentrations These data could be useful both here and in simpler studies of single-compartment bacteria even in the analysis of species but not much information is available. 

Finally, note that eukaryote chemotypes have as a general feature the increase inside the cell vesicles of elements, here calcium, previously confined to the outside of prokaryotes, but these increases are different in different vesicles and organelles and in different organisms separating cells into different chemotypes. This is also seen in their minerals. There is as yet far too little quantitative analysis of calcium or indeed of elements generally to allow us to build a full picture of chemotypes together with genotypes (see Table 8.22). 

Kingsman, S.M. and Kingsman, AJ. (1988). Genetic Engineering An Introduction to Gene Analysis and Exploitation in Eukaryotes, Blackwell Scientific Publications, Oxford, UK, pp 522. 

There are numerous protocols for polysomal gradients preparations that differ mainly at the step for harvesting the cells, and the gradient composition and separation times. The protocol presented later was optimized for isolation of polysomal mRNA from the yeast Saccharomyces cerevisiae, yet many steps will be similar to other eukaryotes and the procedure can easily be modified for other organisms. We will use this protocol as a template on which we will indicate and highlight points that are critical for the microarray analysis. Generally, the RNA isolated by this protocol can be used for analysis by DNA microarray, Northern blot, or RT-PCR. 

Fig. 10.2. FSPIM analysis of the interaction between maize transcriptional coactivators—GCN5 and ADA2—fused to CFP and YFP. GCN5 is a histone acetyltransferase that, in conjunction with adaptor protein ADA2, modulates transcription in diverse eukaryotes by affecting the acetylation status of the core histones in nucleosomes [63]. CFP- and YFP-tagged proteins, expressed in protoplasts, were excited by the 458 nm and the 514 nm laser lines sequentially. CFP fluorescence was selectively detected by an FIFT 458 dichroic mirror and BP 470-500 band pass emission filter while YFP fluorescence was selectively detected by using an HFT 514 dichroic mirror and...

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