An Alcaligenes sp. BR60, isolated from surface runoff waters of the Hyde Park industrial landfill, contained a novel 85 kb catabolic plasmid (pBR60) functional in 3-chlorobenzoate (3Cba) degradation. The plasmid exhibited a spontaneous 3.2% frequency of deletion of a 14 kb fragment specifying 3Cba degradation. The deletion mutant BR 40 and mitomycin C cured strains were not able to grow on 3Cba and had reversion frequencies of less than 10(-10) cell-1 generation-1. Transformation or conjugation of pBR60 into cured strains restored catabolic activity. An EcoRI, BglII, HindIII and SalI restriction map of the deletion region was constructed, and EcoRI and HindIII fragments spanning the deletion region of the plasmid were cloned in pUC18. Conjugation of resistance plasmid R68.45 into Alcaligenes sp. BR 60, with selection on antibiotics, resulted in the elimination of pBR60 and maintenance of unaltered R68.45. In 30% of the exconjugants, 3Cba degradative capacity was retained, although variation in the regulation of 3Cba degradation was observed in these strains. Hybridization of deletion region fragments to BglII digested total DNA of BR60 and the R68.45 cured exconjugants revealed the presence of pBR60 deletion region sequences in the chromosome of exconjugants. Hybridization also revealed a repeated sequence flanking the deletion region of pBR60. Selection on 4-chlorobenzoate as a sole source of carbon and energy resulted in the isolation of 4Cba+ mutants of Alcaligenes sp. BR60.

The parDE2 operon of Vibrio cholerae encodes a type II TA system, which is one of three loci in the superintegron of small chromosome II that show modest similarity to the parDE operon of plasmid RK2. ParE2, like plasmid RK2-encoded ParE, inhibits DNA gyrase, an essential topoisomerase that is also the target of quinolone antibacterial agents. Mechanistic understanding on ParE2 toxin inhibition by direct interaction with its cognate antitoxin and transcriptional autoregulation of the TA system are currently lacking. ParD2, the ribbon-helix-helix (RHH) antitoxin, auto-represses the parDE2 promoter. This repression is enhanced by ParE2, which therefore functions as a transcriptional co-repressor. Here we present protein-DNA interaction studies and high-resolution X-ray structures of the ParD2:ParE2 complex and isolated ParD2 antitoxin, revealing the basis of toxin inhibition and autoregulation of the TA operon by conditional cooperativity. Native mass spectrometry, SAXS and MALS studies confirm the presence of different oligomerization states of ParD2 in solution and the role of the DNA-binding hexameric ParD26:ParE22 assembly in transcriptional repression.

In India antibiotics are frequently used for preventing and controlling bacterial pathogens in carp aquaculture system, yet no studies have been performed to evaluate the ecological impact of its intensive and prolonged use. In this work the frequency of oxytetracycline-resistant bacteria from water, palletized feed and different life stages of fish from Indian freshwater carp aquaculture system as well as the level of resistance of selected strains was investigated. Viable as well as antibiotic-resistant bacterial counts were performed by spread plate method in culture media supplemented with the oxytetracycline. Sixty two resistant Gram negative isolates which represented the oxytetracycline-resistant bacterial population, were randomly selected on nutrient agar supplemented with oxytetracycline (50µg/ml) from carp farms and feed pellet samples. Among these bacterial isolates Flavobacterium (21%), Alcaligenes (14.5%), Aeromonas (11%), Pseudomonas (10%) and Enterobacteriace (19%) were the most frequent. The Escherichia, Serratia, Citrobacter, Enterobacter, Shigella and Proteus from Enterobacteriace were recovered. Twelve isolates of oxytetracycline resistant bacteria were mainly dominated in adult fishes by the genus Flavobacterium (23%) and Enterobacteriace(41%). Selected strains exhibited high levels of oxytetracycline resistance with minimum inhibitory concentrations (MICs) ranging from 50 to 600µg/ml.This study shows the presence of an important population of oxytetracycline-resistant bacteria in the microflora of Indian carp aquaculture farms. Therefore the environment of these farms might play important roles as reservoirs of bacteria carrying genetic determinants for high level tetracycline resistance, prompting an important risk to public health.

Musa accuminata is the member of family Musaceae. The fruits of M. accuminata are harvested in unriped green in order to better withstand transportation and to slow down the natural ripening process because of several weeks travel from the production areas to the end markets. The ripening process is a multi step reaction involves a number of complex enzymes. The present work is aimed to computational characterization of these ripening enzymes in order to understand the ripening mechanism. In this work identification of the enzymes/proteins involved in ripening process are characterized by their conserved domains (conserved, 3-dimension region). The domains are identified by using the computational tool, CDART (Conserved Domain Architecture Retrieval Tool). By using this applied approach a number of conserved domains were identified, out of these enzymes, the domain pattern of Aspartate aminotransferase (AAT_Like), Glycosyl hydrolase (Glyco_Hydr) and oxoglutarate (20G_Fe) were reported the most conserved followed by Ethylene insensitive (EIN3), GT1, DPBB & Pollen in ripening enzyme complexes. The data, thus, obtained provide new insights in order to understand the role of the proteins playing in different stages of ripening process. These results provide a basis for further studies on the molecular mechanism of ripening process and food technological applications for delayed ripening in fruits, where ripening proteins plays an important role in.