ABSTRACT
Recombinant Gluconacetobacter diazotrophicus containing Cry1Ac gene from Bacillus thuringiensis var. kurstaki borne on pKT230, shuttle vector, was generated. PCR amplification of Cry1Ac gene present in recombinant G. diazotrophicus yielded a 278-bp DNA product. The nitrogenase assay has revealed that the recombinant G. diazotrophicus in sugarcane stem produced similar levels of nitrogenase compared to wild-type G. diazotrophicus. The presence of 130-kDa protein in apoplastic fluid from sugarcane stem harvested from pots inoculated with recombinant G. diazotrophicus shows that the translocated G. diazotrophicus produces 130-kDa protein which is recognized by the hyperimmune antiserum raised against 130-kDa protein. The first instar Eldana saccharina neonate larvae that fed on artificial medium containing recombinant G. diazotrophicus died within 72 h after incubation.
Subject(s)
Bacillus thuringiensis/genetics , Bacterial Proteins , Endotoxins , Gluconacetobacter/genetics , Hemolysin Proteins , Nitrogen Fixation , Pest Control, Biological , Recombination, Genetic , Animals , Bacillus thuringiensis/metabolism , Bacillus thuringiensis Toxins , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Electrophoresis, Polyacrylamide Gel , Endotoxins/genetics , Endotoxins/metabolism , Gluconacetobacter/metabolism , Hemolysin Proteins/genetics , Hemolysin Proteins/metabolism , Immunoblotting , India , Larva/microbiology , Lepidoptera/growth & development , Lepidoptera/microbiology , Microscopy, Electron, Scanning , Nitrogenase/genetics , Nitrogenase/metabolism , Polymerase Chain Reaction , Saccharum/metabolism , Saccharum/microbiologyABSTRACT
2,2'-bipyridyl based copper complex I: [CuC24H22N6O10] at 10 nM and complex Ia: [Cu2C32H43N8O3](PF6)4 at 7 nM exhibited 50% inhibition of lymphocyte proliferation and less than 20% cytotoxicity in peripheral blood mononuclear cells (PBMCs). Further, pro-inflammatory cytokines such as TNF-alpha and IL-1beta and pro-inflammatory mediator such as Nitric Oxide (NO) besides inducible Nitric Oxide Synthase (iNOS) were inhibited by the copper complexes. They also down regulated the expression of cyclooxegenase-2 (COX-2), yet another mediator of inflammation. Immunoblot analysis revealed the inhibitory effect of these complexes on phosphorylated forms protein kinases (MAPKs) such as ERK1/2, JNK, and p38 in mitogen induced PBMCs.
