Broad-host-range vector system for synthetic biology and biotechnology in cyanobacteria

Inspired by the developments of synthetic biology and the need for improved genetic tools to exploit cyanobacteria for the production of renewable bioproducts, we developed a versatile platform for the construction of broad-host-range vector systems. This platform includes the following features: (i) an efficient assembly strategy in which modules released from 3 to 4 donor plasmids or produced by polymerase chain reaction are assembled by isothermal assembly guided by short GC-rich overlap sequences. (ii) A growing library of molecular devices categorized in three major groups: (a) replication and chromosomal integration; (b) antibiotic resistance; (c) functional modules. These modules can be assembled in different combinations to construct a variety of autonomously replicating plasmids and suicide plasmids for gene knockout and knockin. (iii) A web service, the CYANO-VECTOR assembly portal, which was built to organize the various modules, facilitate the in silico construction of plasmids, and encourage the use of this system. This work also resulted in the construction of an improved broad-host-range replicon derived from RSF1010, which replicates in several phylogenetically distinct strains including a new experimental model strain Synechocystis sp. WHSyn, and the characterization of nine antibiotic cassettes, four reporter genes, four promoters, and a ribozyme-based insulator in several diverse cyanobacterial strains.     

Effect of environmental factors on disease development caused by the fungal pathogen Plectosporium alismatis on the floating-leaf stage of starfruit (Damasonium minus), a weed of rice

Starfruit, Damasonium minus (R. Br.) Buch, is a herbicide-resistant, aquatic-weed occurring in Australian rice fields. The fungal pathogen Plectosporium alismatis (Oudem) W.M. Pitt, W. Gams & U. Braun is a potential mycoherbicide for the control of this weed. Previous work showed that the fungus can suppress the growth of starfruit juvenile plants. This paper reports the effect of environmental factors on the disease development on starfruit at the floating-leaf stage. Fungal inoculation of plants in advanced phases of floating leaf stage reduced significantly (P<0.05) the green leaf area, but only when the percentages of diseased areas were deducted. However, inoculation when the first floating leaf was emerging reduced significantly (P<0.001) both, plant dry above-ground biomass and leaf area.     

Expression of ERBB gene family in females with breast cancer and its correlation with clinicopathological characteristics of the disease

Molecular Biology Reports - Breast cancer (BC) is the most prevalent and fatal cancer in women. Given that there are very few studies investigating the overexpression of four members of ERBB genes,...     

Design of novel granulopoietic proteins by topological rescaffolding

Computational protein design is rapidly becoming more powerful, and improving the accuracy of computational methods would greatly streamline protein engineering by eliminating the need for empirical optimization in the laboratory. In this work, we set out to design novel granulopoietic agents using a rescaffolding strategy with the goal of achieving simpler and more stable proteins. All of the 4 experimentally tested designs were folded, monomeric, and stable, while the 2 determined structures agreed with the design models within less than 2.5 Å. Despite the lack of significant topological or sequence similarity to their natural granulopoietic counterpart, 2 designs bound to the granulocyte colony-stimulating factor (G-CSF) receptor and exhibited potent, but delayed, in vitro proliferative activity in a G-CSF-dependent cell line. Interestingly, the designs also induced proliferation and differentiation of primary human hematopoietic stem cells into mature granulocytes, highlighting the utility of our approach to develop highly active therapeutic leads purely based on computational design.

De novo designed cytokines that activate the G-CSF receptor show that the receptor-binding information can be encoded onto stable, miniaturised protein scaffolds that possess potent granulopoietic activity; such novel proteins provide for ideal candidates for protein-based therapeutics.     

Seasonality of the endogenous activity rhythm in Talitrus saltator (Montagu) from a sandy beach in northeastern Tunisia

The endogenous activity of the sandhopper Talitrus saltator (Montagu) has been investigated in a population from a relatively exposed beach in northeast Tunisia. The animals were found to exhibit a nocturnal circadian rhythm of locomotor activity, with no evidence of a circatidal component. The rhythm was monitored in individual animals over the four seasons, under constant laboratory conditions and seasonal variation in free-running period; the signal-to-noise ratio and waveform of the rhythm have been investigated, as well as the incidence of rhythmic animals in the population. The variation in these rhythm parameters is considered in relation to changes in environmental conditions prevailing at the site of collection.     

Evaluation of Efficiency of Modified Polypropylenimine (PPI) with Alkyl Chains as Non-viral Vectors Used in Co-delivery of Doxorubicin and TRAIL Plasmid

In this study, co-delivery system was achieved via plasmid encoding TNF related apoptosis inducing ligand (pTRAIL) and doxorubicin (DOX) using carrier based on polypropylenimine (PPI) modified with 10-bromodecanoic acid. Incorporation of alkylcarboxylate chain to PPIs (G4 and G5) could improve transfection efficiency via overcoming the plasma membrane barrier of the cells and decrease cytotoxicity of PPI. Characterization of fabricated NPs revealed that PPI G5 in which 30% of primary amines were substituted by alkyl carboxylate chain (PPI G5-Alkyl 30%) has higher drug loading as compared to the other formulations. PPI G5-Alkyl 30% indicated a decreased drug release may be due to alkyl chains on the surface of PPI, which serve as an additional hindrance for drug diffusion. In vitro cytotoxicity experiments demonstrated that co-delivery system induced apoptosis of tumor cells more efficiently than each of delivery system alone. Furthermore, these results revealed that our combined delivery platform of pTRAIL and DOX using Alkyl-modified PPI G5 can significantly improve the anti-tumor activity and this strategy might develop a new therapeutic window for cancer treatment.