RRC ID 33480
著者 Rondahl H, Nilsson B, Holmgren E.
タイトル Fusions to the 5' end of a gene encoding a two-domain analogue of staphylococcal protein A.
ジャーナル J Biotechnol
Abstract A novel gene fusion system has been constructed for fusions to the 5' end of gene zz, encoding a two-domain analogue of staphylococcal protein A designated ZZ. Four different genes were fused to the 5' end of zz, and their gene products were analyzed. One of the genes encodes a protein located intracellularly in Escherichia coli and the other three genes encode gene products destined for secretion across the cytoplasmic membrane by the presence of an amino terminal signal sequence. After production in E. coli, the fusion proteins were purified in a single step by IgG-affinity chromatography. The purified ZZ fusions could be used directly for amino terminal sequencing to confirm the start of translation of the intracellular product and the processing of the signal peptide of the translocated products. This is the first example of ZZ fusions to the C-terminus of gene products. To simplify the general use of fusions to the 5' end of zz, a new plasmid vector was constructed containing a multi restriction enzyme cloning linker and the lacZ' gene which enables screening for production in alpha-complementing supE strains of E. coli on indicator plates.
巻・号 25(3)
ページ 269-87
公開日 1992-9-1
DOI 10.1016/0168-1656(92)90161-2
PII 0168-1656(92)90161-2
PMID 1368804
MeSH Amino Acid Sequence Base Sequence Cloning, Molecular* Escherichia coli / genetics Molecular Sequence Data Plasmids Protein Sorting Signals / analysis Recombinant Fusion Proteins / biosynthesis Recombinant Fusion Proteins / isolation & purification Staphylococcal Protein A / biosynthesis Staphylococcal Protein A / genetics* Staphylococcal Protein A / isolation & purification
IF 3.503
引用数 13
WOS 分野 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
リソース情報
原核生物(大腸菌) pKP497