A plasmid-based expression system to study protein–protein interactions at the Golgi in vivo

There is still an unmet need for simple methods to verify, visualize, and confirm protein–protein interactions in vivo. Here we describe a plasmid-based system to study such interactions. The system is based on the transmembrane domain (TMD) of the EF-hand Ca²⁺ sensor protein calneuron-2. We show that fusion of 28 amino acids that include the TMD of calneuron-2 to proteins of interest results in prominent localization on the cytoplasmic side of the Golgi. The recruitment of binding partners to the protein of interest fused to this sequence can then be easily visualized by fluorescent tags.     

Influence of root and leaf traits on the uptake of nutrients in cover crops

Plant and Soil - The objectives of this study were to determine the spatial structure of soil respiration (Rs) in a naturally-regenerated longleaf pine forest and to assess the ecological factors...     

The sexual phase of the diatom <Emphasis Type="Italic">Pseudo-nitzschia multistriata</Emphasis>: cytological and time-lapse cinematography characterization

Pseudo-nitzschia is a thoroughly studied pennate diatom genus for ecological and biological reasons. Many species in this genus, including Pseudo-nitzschia multistriata, can produce domoic acid, a toxin responsible for amnesic shellfish poisoning. Physiological, phylogenetic and biological features of P. multistriata were studied extensively in the past. Life cycle stages, including the sexual phase, fundamental in diatoms to restore the maximum cell size and avoid miniaturization to death, have been well described for this species. P. multistriata is heterothallic; sexual reproduction is induced when strains of opposite mating type are mixed, and proceeds with cells producing two functionally anisogamous gametes each; however, detailed cytological information for this process is missing. By means of confocal laser scanning microscopy and nuclear staining, we followed the nuclear fate during meiosis, and using time-lapse cinematography, we timed every step of the sexual reproduction process from mate pairing to initial cell hatching. The present paper depicts cytological aspects during gametogenesis in P. multistriata, shedding light on the chloroplast behaviour during sexual reproduction, finely describing the timing of the sexual phases and providing reference data for further studies on the molecular control of this fundamental process.     

Die Immunsysteme der Bakterien

The immune systems of bacteria and important applications in biotechnology and medicine At the end of the 70s of the last century, a new technique has been developed allowing the synthesis of genes and the inducible expression of their recombinant proteins using restriction enzymes and vectors, mainly plasmids. This era has been designated as genetic engineering and is being replenished by the CRISPR‐Cas9 technology know as genome editing. This technology is about to revolutionize alterations in the genomes of all types of organisms, including bacteria, fungi, plants, animals and even humans. It allows the introduction and elimination of point mutations and even whole genes in all organisms. Important goals are the genetic optimization of crop plants and animals, fighting against cancer in humans and elimination of human viruses and pathogenic multi‐resistant bacteria. Important drawbacks are OFF‐targets which can cause mutations in any gene or influence their expression.