PubMed · 2575205
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J J Villafranca. 1989. Paramagnetic probes of macromolecules.. https://doi.org/10.1016/0076-6879(89)77023-3
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The impact of three different RNA isolation methods on the community analysis of metabolically active bacteria was determined by reverse transcription (RT) and PCR amplification of 16S rRNA genes and subsequent terminal restriction fragment length polymorphism (T-RFLP) analysis. Furthermore, soil samples were stored at different conditions in order to evaluate the effect of soil conservation methods on the outcome of the population analysis. The quality of mRNA was assessed by reverse transcription and PCR amplification of eubacterial glutamine synthetase genes. Our results indicated that the community composition as well as the abundance of individual members were affected by the kind of RNA isolation method. Furthermore, the extraction method influenced the recovery of mRNA. Lyophilization, storage at -20 degrees C as well as storage in glycerol stocks at -80 degrees C proved to be equally appropriate for the storage of soils and subsequent RNA isolation.
Glutamine synthetase from Streptomyces lincolnensis was purified to electrophoretic homogeneity. The effects of pH, temperature and metal ions on the purified enzyme were studied, and the substrate specificity of glutamine synthetase were reported. Under the standard biosynthetic assay system, the substrate saturation curves of glutamine synthetase from Streptomyces lincolnensis for ammonia did not agree with the Michaelis-Menten relationship, with either Mn2+ or Mg2+ as divalent cation, and Hill constants not equal to 1. The K(m) values of GS for Glu and ATP were different with different divalent cation.
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