Expression and purification of the recombinant ConBr (Canavalia brasiliensis lectin) produced in Escherichia coli cells.
ConBr, a D-glucose/D-mannose-specific lectin from Canavalia brasiliensis seeds, was produced in Escherichia coli from a (c)DNA clone subcloned to pET15b expression vector. The recombinant lectin (rConBr) was purified by one-step immobilized metal-affinity chromatography using an amino-terminal hexahistidine tag. By SDS-PAGE and Western blot, rConBr was highly pure with an apparent molecular mass of 37 kDa. N-terminal sequence analysis revealed a single sequence, confirming the identity of the expressed protein as the pre-pro-ConBr.
Depto. de Analises Clínicas e Toxicológicas, Universidade Federal do Ceara, Ceara, Brasil.
Nadia A P
NA
Nogueira
Moema B
MB
Grangeiro
Rodrigo M S
RM
da Cunha
Marcio V
MV
Ramos
Maria A O
MA
Alves
Edson H
EH
Teixeira
Manoel
M
Barral-Netto
Juan J
JJ
Calvete
Benildo S
BS
Cavada
Thalles B
TB
Grangeiro
D015153:Blotting, Western; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D007887:Fabaceae / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D006639:Histidine / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D037102:Lectins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D009532:Nickel / Q000737:chemistry / Q000378:metabolism; D037121:Plant Lectins; D010957:Plasmids; D011994:Recombinant Proteins
D037102:Lectins; D037121:Plant Lectins; D011994:Recombinant Proteins; D006639:Histidine; D009532:Nickel