Towards a Systems Approach in the Genetic Analysis of Archaea: Accelerating Mutant Construction and Phenotypic Analysis in Haloferax volcanii

Figure 1

Generation of H. volcanii gene deletion constructs. Regions of approximately 600 bp flanking the gene of interest were amplified from the H. volcanii chromosome (a) Regions of complementarity between primers are ^' and ^”. Complementarity sequences were 15 b, as follows: NF: CGGGCCCCCCCTCGAG; NR: GACGCGTTCATATGC; CF: GCATATGAACGCGTC CR: CGGGCTGCAGGAATTC and were designed to yield unique MluI NdeI sites. Plasmid pTA131 was digested with EcoRI and XhoI (c) and the three fragments recombined using In-Fusion (d). Refer to Materials and Methods for details of the techniques used.