Heterologous expression of bacteriocin E-760 in Chlamydomonas reinhardtii and functional analysis

The use of antimicrobial peptides (AMPs) synthesized by bacteria (bacteriocins) is an alternative for combating multidrug resistant bacterial strains and their production by recombinant route is a viable option for their mass production. The bacteriocin E-760 isolated from the genus Enterococcus sp. has been shown to possess inhibitory activity against Gram-negative and Gram-positive bacteria. In this study, the expression of a chimeric protein coding for E-760 in the nucleus of C. reinhardtii was evaluated, as well as, its antibacterial activity. The synthetic gene E-760S was inserted into the genome of C. reinhardtii using Agrobacterium tumefaciens. A transgenic line was identified in TAP medium with hygromycin and also by PCR. The increment in the culture medium temperature of the transgenic strain at 35 °C for 10 minutes, increased the production level of the recombinant protein from 0.14 (Noninduced culture, NIC) to 0.36% (Induced culture, IC) of total soluble proteins (TSP); this was quantified by an ELISA assay. Recombinant E-760 possesses activity against Staphylococcus aureus in 0.34 U log, Streptococcus agalactiae in 0.48 U log, Enterococcus faecium in 0.36 U log, Pseudomonas aeruginosa in 2 U log and for Klebsiella pneumoniae, the activity was 0.07 U log. These results demonstrate that the nucleus transformation of C. reinhardtii can function as a stable expression platform for the production of the synthetic gene E-760 and it can potentially be used as an antibacterial agent.     

Evidence from bioinformatics, expression and inhibition studies of phosphoinositide-3 kinase signalling in Giardia intestinalis

Background  

Giardia intestinalis is a parasitic protozoan and major cause of diarrhoeal disease. Disease transmission is dependent on the ability of the parasite to differentiate back and forth between an intestine-colonising trophozoite and an environmentally-resistant infective cyst. Our current understanding of the intracellular signalling mechanisms that regulate parasite replication and differentiation is limited, yet such information could suggest new methods of disease control. Phosphoinositide-3 kinase (PI3K) signalling pathways have a central involvement in many vital eukaryotic processes, such as regulation of cell growth, intracellular membrane trafficking and cell motility. Here we present evidence for the existence of functional PI3K intracellular signalling pathways in G. intestinalis.     

Inhibiteurs non acides et dormance embryonnaire chez Taxus baccata

Non-acidic inhibitors and embryo dormancy in Taxus baccata L. Embryo dormancy of Taxus baccata is eliminated when the embryos are continuously kept in sterile nutritive liquid medium. After 3 weeks of culture, an important non-acidic inhibitory complex can be extracted from this liquid medium. At least three substances are involved: two pigments and a compound with some properties that suggest xanthoxin. These substances are neither found in embryos taken directly from the seeds, nor in liquid medium after 8 days of culture, which is the time necessary and sufficient to allow germination after transfer on agar medium. Such behaviour is quite different from that of ABA previously studied and indicates that these non-acidic inhibitors appear late during the culture and are not directly involved in embryo dormancy.     

c-AMP and Morphogenesis of Acetabularia

The c-AMP content has been found to double when Acetabularia develop from 5–10 mm long to grown or almost full-grown algae.
The biological significance of this fact has been approached by studying the effects of drugs known to influence the intracellular c-AMP content on the development of Acetabularia. When grown in the presence of theophyllin or papaverin, inhibitors of phosphodiesterase, the Acetabularia display a striking response during the exponential growth period; the final length, however, is not affected. Both substances increase the c-AMP content of the algea. Isoproterenol, which activates adenylate cyclase in many systems, also influences Acetabularia during the exponential growth period and, in addition, slightly affects cap formation.
The change in c-AMP content in the course of development and the effects of drugs influencing (theophyllin and papaverin) or likely to influence (isoproterenol) the c-AMP content of the algae suggest that this nucleotide plays a role at the time of intense growth.
The same phosphodiesterase activity has been found in the 5–10 mm and the 19–25 mm long algae, whereas two enzymes were found in cap-bearing Acetabularia.
The results are discussed as well as the involvement of c-AMP in the development of this alga.