Article Title: Expression, purification and crystallization of CTB-MPR, a candidate mucosal vaccine component against HIV-1
Figure Lengend Snippet: ( a ) The architecture of gp41. FP (residues 512–527), fusion peptide; FPPR (residues 528–539), fusion peptide proximal region; NHR (residues 540–590), N-terminal heptad-repeat region; CHR (residues 628–661), C-terminal heptad-repeat region; MPER (residues 662–684), membrane-proximal external region; MPR (residues 647–684, hatched), membrane-proximal region; TM (residues 685–705), transmembrane domain; CTD (residues 706–856), cytoplasmic C-terminal domain. ( b , c , d ) DNA constructs for the expression in E. coli of the indicated CTB-MPR fusion proteins are based on elements of the pET-22b expression vector. P, T7 bacteriophage promoter; 5′-UTR, upstream untranslated region; pelB, the periplasmic targeting sequence of pectate lyase B of Erwinia carotovora ; CTB, cholera toxin B subunit; MPR, the membrane-proximal region of the gp41 protein of HIV-1; 3′-UTR, downstream untranslated region; T, T7 terminator. The GPGP and AAAA linkers are indicated above their respective constructs. The three constructs encode the fusion proteins CTB GPGP MPR ( b ), CTBMPR ( c ) and CTB AAAA MPR ( d ) with expected molecular masses (after the processing of the pelB leader sequence) of 16.7, 16.4 and 16.7 kDa, respectively.
Article Snippet: Vectors for bacterial expression of CTB-MPR fusion protein variants The expression vectors used in this study were all based on the Escherichia coli periplasmic targeting vector pET-22b(−) (Novagen; Figs. 1 b , 1 c and 1 d ).
Techniques: Construct, Expressing, CtB Assay, Positron Emission Tomography, Plasmid Preparation, Sequencing