Transferring foreign genes into mushroom mediated by Agrobacterium tumefaciens is a standard technique in genetic engineering. Recombinant human insulin has been greatly used in the treatment of type I diabetes. The production of edible mushroom derived insulin should facilitate oral delivery. In this study we used the Agrobacterium tumefaciens mediated transformation method for the transfer an...

As aerobic chemoorganotrophs, most Agrobacterium strains will grow on a wide range of complex and defined media. Methods commonly used for the culture and storage of other chemoorganotrophs will usually work for agrobacteria as well. Problems with culture or strain maintenance will occur more frequently because of careless technique than because of strain difficulties. Here we describe a few of...

We constructed the broad-host-range plasmid pUCD800 containing the sacB gene of Bacillus subtilis for use in the positive selection and isolation of insertion sequence (IS) elements in gram-negative bacteria. Cells containing pUCD800 do not grow on medium containing 5% sucrose unless the sacB gene is inactivated. By using pUCD800, we isolated a 1.4-kilobase putative IS element from Agrobacteriu...

E Keshamma, B. Madhusudhan, Department of Biochemistry, Center for Nano Science and Technology Research, Kuvenpu University, P.G. Centre, Shivagangotri Davangere 577 002, India; S. Rohini, M. Udaya Kumar*, Department of Crop Physiology, University of Agricultural Sciences, GKVK Bangalore 560 065, India; K. S. Rao, Centre for Ecological Sciences, Indian Institute of Science, Bangalore 560012, In...

In the present study high efficiency Agrobacterium tumefaciens mediated transformation system of wheat (Triticum aestivum L.) cv. Inqilab-91 was developed by exploiting bacterial culture density and acetosyringone concentration. Agrobacterium strain EHA101 harboring binary vector pIG121Hm, containing gene for GUS activity and hygromycin resistance was used in transformation experiments. Differe...

A procedure to enrich for auxotrophic and fermentation mutants of Agrobacterium tumefaciens is described. The method is based on the amplification of the killing power of carbenicillin by the addition of lysozyme. Isolation frequencies of some types of mutants are presented, with and without the application of the proposed procedure. The yield of mutants is usually enhanced a hundredfold per en...

An assay to determine the relative virulence of wild-type and mutant strains of Agrobacterium tumefaciens on leaves of Bryophyllum daigremontiana has been developed. This assay is reproducible, is easy to learn, is not time consuming, and requires little space. The relative virulence of cellulose-minus mutants of A. tumefaciens was investigated with this assay. Some of these mutants were unalte...

Bacteria use conjugation systems, a subfamily of the type IV secretion systems, to transfer DNA to recipient cells. Despite 50 years of research, the architecture and mechanism of action of the channel mediating DNA transfer across the bacterial cell envelope remains obscure. By use of a sensitive, quantifiable assay termed transfer DNA immunoprecipitation (TrIP), we identify contacts between a...