1. A procedure is described for the isolation of intact polyribosomes from the cytoplasm, chloroplasts and mitochondria of Euglena gracilis. 2. All three polyribosomal preparations incorporated labelled amino acids in a system in vitro. The cytoplasmic system was inhibited by chcloheximide but not by chloramphenicol. Both the chloroplast and the mitochondrial systems, however, were inhibited by chloramphenicol but not by cycloheximide. It is shown that mitochondrial polyribosomes, like the polyribosomes from cytoplasm and chloroplasts, can participate directly in protein synthesis without supplementary mRNA being added to the synthesizing system, as in previously reported instances. 3. Sedimentation coefficients were measured for the ribosomes, ribosomal subunits, and rRNA of the cytoplasm, chloroplasts and mitochondria. 4. The G+C content was 55% for cytoplasmic rRNA, 50% for chloroplast rRNA, and 29% for mitochondrial rRNA. 5. The cytoplasmic ribosomal subunits contained a ribonuclease activity that was inhibited by heparin.

Isolation of active polyribosomes from the cytoplasm, mitochondria and chloroplasts of Euglena gracilis.

Publisher Summary This chapter describes physical and chemical methods for disintegrating microbes for preparing biologically active fractions. Direct measurement of cell disintegration can be sometimes dispensed with and the criteria of yield of the desired enzyme activity be relied on, especially when one of the chemical methods of extraction is used. In any comparative work and where control or location is studied by means of a physical method of disruption or with spheroplasts, some method of assessing disintegration is essential. Estimation of cell disintegration by measuring the release of typical cell components can be made quantitatively more precise than direct counting. There is some difficulty in interpretation and in obtaining a normal zero time and 100% standards. For instance, the measurement of the release of soluble protein is often used. This depends partly on the degree of disruption, but also on the method used to separate soluble protein from insoluble debris.

Chapter I The Disintegration of Micro-organisms

Isolation and characterization of DNA from the mitochondrial fraction of Euglena.

Electron transport systems and mitochondrial DNA in trypanosomatidae: A review

A 73 kbp circular DNA was isolated from the colourless euglenoid flagellate Astasia longa. Restriction sites of 12 restriction endonucleases were mapped on this DNA. Southern hybridization using plastid gene probes from Euglena, spinach and tobacco revealed sequence homologies to the genes coding for 16S and 23S ribosomal RNAs, elongation factor Tu (tufA) and the large subunit of ribulose-1,5-bisphosphate carboxylase (rbcL). The locations of these sequences on the restriction map were determined. Sequences homologous to chloroplast genes psaA, psbA, psbD, psbE and atpA are not present. The ribosomal RNA genes are organized in three tandem repeats, each containing one 23S and one 16S rRNA gene. In addition, there is one extra 16S rRNA gene. These results indicate the presence of a truncated form of a plastid DNA in Astasia.

A circular 73 kb DNA from the colourless flagellate Astasia longa that resembles the chloroplast DNA of Euglena: restriction and gene map

Isolation of mitochondria from euglena gracilis.

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Isolation of mitochondria from euglena gracilis.