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has been cited by the following article:

• TITLE: Characterization of Avirulent TnphoA Mutants in Agrobacterium tumefaciens to Enhance Transformation Efficiency

  AUTHORS: Dilip K. Das, Eugene W. Nester

  KEYWORDS: Transposon, Mutagenesis, Agrobacterium tumefaciense, Bradyrhizobium japonicum

  JOURNAL NAME: Advances in Microbiology, Vol.4 No.9, July 28, 2014

  ABSTRACT: Protein fusion with the Escherichia coli alkaline phosphatase is used extensively for the analysis of the topology of membrane protein. Agrobacterium strain A6007 was mutagenized with E. coli strain mm294A plasmid pRK609 having TnphoA to obtain mutants defective in virulence. Because alkaline phosphatase activity is only detected when the PhoA gene product from the transposon is secreted out of the protoplasm, the virulence mutants are located in genes that code for transmembrane or periplasmic proteins. Attempts were made to obtain the sequences adjacent to the TnphoA inserts through several different approaches including Inverse PCR, Cloning, and Tail PCR. Transposon-adjacent sequence was obtained from one membrane anchor subunit in Bradyrhizobium japonicum i.e. succinate dehydrogenase which has enhanced transformation efficiency.