Expression and purification of the recombinant ConBr (Canavalia brasiliensis lectin) produced in Escherichia coli cells.

作者信息

Nogueira Nadia A P, Grangeiro Moema B, da Cunha Rodrigo M S, Ramos Marcio V, Alves Maria A O, Teixeira Edson H, Barral-Netto Manoel, Calvete Juan J, Cavada Benildo S, Grangeiro Thalles B

机构信息

Depto. de Analises Clínicas e Toxicológicas, Universidade Federal do Ceara, Ceara, Brasil.

出版信息

Protein Pept Lett. 2002 Feb;9(1):59-66. doi: 10.2174/0929866023408968.

DOI:10.2174/0929866023408968

PMID:12141925

Abstract

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.

摘要