Preparation and analysis of isogenic mutants in the transferrin receptor protein genes, tbpA and tbpB, from Neisseria meningitidis

作者：Sean Wyndham Irwin, Nuzhat Averil, Cindy Y. Cheng, Anthony Bernard Schryvers · 发表于：Molecular Microbiology · 年份：1993 · DOI：10.1111/j.1365-2958.1993.tb01657.x · 被引用次数：109 · 研究领域：Legionella and Acanthamoeba research、Bacterial Genetics and Biotechnology、Parasitic Infections and Diagnostics

Isogenic mutants were constructed in the tbpA and tbpB genes from Neisseria meningitidis strain B16B6, which code for the transferrin receptor proteins, Tbp1 and Tbp2. Insertion mutants of the tbpA and tbpB genes were obtained by shuttle mutagenesis and by in vitro cassette mutagenesis, respectively. The isogenic mutants were verified by Southern blot and Western blot analysis. Isogenic mutants deficient in Tbp1 or Tbp2 demonstrated a reduced transferrin binding activity in intact cells and total membranes but were incapable of utilizing transferrin iron for growth. Tbp1 could be isolated by affinity methods from the mutant lacking Tbp2 but isolation of Tbp2 from the mutant lacking Tbp1 required the presence of exogenous Tbp1.