Cloning and characterization of the Neisseria meningitidis asd gene

Lee Ann Hatten, Herbert P. Schweizer, Nuzhat Averill, Ling Wang, Anthony B. Schryvers

Research output: Contribution to journalArticlepeer-review

9 Scopus citations


The asd mutants of Gram- and some Gram+ bacteria have an obligate requirement for diaminopimelic acid (DAP), an essential constituent of the cell wall of these organisms. In environments deprived of DAP, i.e., mammalian tissues, they will undergo lysis. This has previously been exploited to develop vaccine strains of Salmonella typhimurium and Streptococcus mutaons. As a first step for the development of a biosafe Neisseria meningitidis laboratory strain, we have cloned the asd from wild-type strain B16B6 by complementation of an Eschirichia coli asd mutant. By subcloning and insertion mutagenesis, the N. meningitidis asd was localized to a 1.5-kb DNA fragment. In a T7 RNA polymerase-T7 promoter expression system, a 38-kDa protein was strongly expressed from this DNA fragment. The N-terminal amino acid (aa) sequence was deduced from the nucleotide sequence, which was determined with the help of an in-frame Asd'::'LacZ protein fusion. A comparison of the N-terminal aa of the Asd proteins from N. meningitidis and E. coli revealed 70% identity, suggesting that the Asd protein may be highly conserved among Gram- bacteria.

Original languageEnglish (US)
Pages (from-to)123-128
Number of pages6
Issue number1
StatePublished - Jul 15 1993
Externally publishedYes


  • Cell wall
  • biosafety
  • gene fusion
  • reading frame
  • recombinant DNA
  • β-galactosides

ASJC Scopus subject areas

  • Genetics


Dive into the research topics of 'Cloning and characterization of the Neisseria meningitidis asd gene'. Together they form a unique fingerprint.

Cite this