Domain organization of NgBR and its
AR A014418 and purification. (A) Bioinformatics characterization reveals that NgBR contains a putative signal sequence (1–46), an ectodomain (47–120), a type 1A transmembrane domain (121–139) and a cytoplasmic domain (140–297). (B) Coomasie brilliant blue stained SDS–PAGE gel showing the expression and affinity-purification of the NgBR ectodomain. Line 1: total cell extract after IPTG induction; line 2: supernatant of the cell lysate; line 3: GST beads with bound GST-fused ectodomain; line 4: GST beads after in-gel thrombin cleavage; line 5: protein molecular weight markers. (C) Coomasie brilliant blue stained SDS–PAGE gel showing the expression and affinity-purification of the NgBR cytoplasmic domain. Line 1: total cell extract before IPTG induction; line 2: total cell extract after IPTG induction; line 3: supernatant of the cell lysate; line 4: pellet of the cell lysate; line 5: Ni-agrose beads with bound cytoplasmic domain in the presence of 8 M urea; line 6: protein molecular weight markers. (D) Coomasie brilliant blue stained SDS–PAGE gel showing the
expression and affinity-purification of the full-length of NgBR. Line 1: total cell extract before IPTG induction; line 2: total cell extract after IPTG induction; line 3: supernatant of the cell lysate; line 4: pellet of the cell lysate; line 5: Ni-agrose beads with bound NgBR in the presence of 8 M urea; line 6: protein molecular weight markers.
