A novel expression vector, designated as pHisJM, for producing recombinant His-fusion proteins

Junko Masuda, Eiji Takayama, Ayano Satoh, Kyoko Kojima-Aikawa, Kimihiro Suzuki, Isamu Matsumoto

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Compared to glutathione S-transferase (GST), tagging with hexahistidine residues (His) has several merits: low levels of toxicity and immunogenicity, a smaller size and no electric charge. We have constructed a novel expression vector, designated as pHisJM (EMBL/GenBank/DDJB accession no. AB116367), for producing recombinant His-fusion proteins. This vector was constructed by replacing GST and multiple cloning site (MCS) cassettes in pGEX-5X-3 with those of hexahistidine and MCS derived from pRSET C vector. Human annexin IV (Anx IV) was used as target protein. His-Anx IV fusion protein was expressed using pHisJM and gave a 40 kDa band when immuno-stained with anti-His mAb or anti-Anx IV mAb as predicted. To compare expression efficiency, a Anx IV cDNA inserted-pHisJM or pGEX-5X-3 was transformed into Escherichia coli DH5α, JM109, BL21 and BL21(DE3). Using pHisJM, Anx IV protein was highly expressed in all cell strains. In addition to the merits of using His-tag, pHisJM has several advantages: 1) it has high expression efficiency; 2) it can be used in any Escherichia coli strain; and 3) it can be used in a single strain of Escherichia coli in all steps from plasmid construction to the expression of the target gene.

Original languageEnglish
Pages (from-to)1543-1548
Number of pages6
JournalBiotechnology Letters
Volume26
Issue number20
DOIs
Publication statusPublished - Oct 2004
Externally publishedYes

Fingerprint

His-His-His-His-His-His
Annexin A4
Fusion reactions
Escherichia coli
Proteins
Cloning
Glutathione Transferase
Electric charge
Organism Cloning
Toxicity
Genes
Nucleic Acid Databases
Plasmids
Complementary DNA

Keywords

  • Escherichia coli expression system
  • Expression vector
  • Hexahistidine tag
  • Protein purification
  • tac promoter

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology
  • Microbiology
  • Bioengineering

Cite this

A novel expression vector, designated as pHisJM, for producing recombinant His-fusion proteins. / Masuda, Junko; Takayama, Eiji; Satoh, Ayano; Kojima-Aikawa, Kyoko; Suzuki, Kimihiro; Matsumoto, Isamu.

In: Biotechnology Letters, Vol. 26, No. 20, 10.2004, p. 1543-1548.

Research output: Contribution to journalArticle

Masuda, Junko ; Takayama, Eiji ; Satoh, Ayano ; Kojima-Aikawa, Kyoko ; Suzuki, Kimihiro ; Matsumoto, Isamu. / A novel expression vector, designated as pHisJM, for producing recombinant His-fusion proteins. In: Biotechnology Letters. 2004 ; Vol. 26, No. 20. pp. 1543-1548.
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