Fusion expression and purification of HA20-lys10 in E. coli and its gel retardation experiments

Xinghui Sun, Yu Qing Zhang, Xia Wang, Min Hou, Li Tan, Ping Li, Yun Song Zhu

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Purpose: To express and purify the fused protein containing 20 residues of amino-terminal of influenza hemagglutinin HA2 and ten lysines, and investigate its ability of binding plasmid. Methods: The fused gene HA20-1ys10 containing gene of 20 residues of amino-terminal of influenza hemagglutinin HA2 and ten lysines was amplified by PCR, and was cloned into prokaryotic expression vector pET32a (+). The recombinant expression plasmid pET-HA-K was transformed into E. coli BL21 (DE3). When A600 reached 0. 6, IPTG was added to induce the expression of recombinant protein. The expressed protein was purified by one-step affinity chromatography, and its ability of binding plasmid was investigated via gel retardation experiments. Results: After induction for 3 hours by 1 mmol/L IPTG, there was a high level expression of recombinant protein, and most of them were soluble. After purification by affinity chromatography, the purity of HA20-1ys10 was up to 85%, and 20 μg HA20-1ys10 could retard plasmid completely. Conclusions: These results indicate that HA20-1ys10 is hopeful to be used in receptor-mediated gene transfer to improve the efficiency.

Original languageEnglish (US)
Pages (from-to)32-35
Number of pages4
JournalFudan University Journal of Medical Sciences
Volume31
Issue number1
StatePublished - Jan 1 2004

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Plasmids
Gels
Escherichia coli
Isopropyl Thiogalactoside
Hemagglutinins
Affinity Chromatography
Recombinant Proteins
Human Influenza
Lysine
Genes
Proteins
Polymerase Chain Reaction

Keywords

  • Expression and purification
  • Fused gene
  • Gel retardation experiment
  • Hemagglutinin

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Fusion expression and purification of HA20-lys10 in E. coli and its gel retardation experiments. / Sun, Xinghui; Zhang, Yu Qing; Wang, Xia; Hou, Min; Tan, Li; Li, Ping; Zhu, Yun Song.

In: Fudan University Journal of Medical Sciences, Vol. 31, No. 1, 01.01.2004, p. 32-35.

Research output: Contribution to journalArticle

Sun, Xinghui ; Zhang, Yu Qing ; Wang, Xia ; Hou, Min ; Tan, Li ; Li, Ping ; Zhu, Yun Song. / Fusion expression and purification of HA20-lys10 in E. coli and its gel retardation experiments. In: Fudan University Journal of Medical Sciences. 2004 ; Vol. 31, No. 1. pp. 32-35.
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AU - Zhu, Yun Song

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