A naturally enhanced green fluorescent protein from magnificent sea anemone (Heteractis magnifica) and its functional analysis

Hongbin Tu, Qian Xiong, Suilan Zhen, Xiaofen Zhong, Lisheng Peng, Huiping Chen, Xiaoyu Jiang, Wenhua Liu, Wenli Yang, Jianwen Wei, Meiling Dong, Wenyan Wu, Anlong Xu

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

A novel fluorescent protein termed hmGFP homologous to the green fluorescent protein (GFP) from Aequorea victoria was cloned from the tentacles of sea anemone Heteractis magnifica by EST sequencing and analysis of cDNA library and followed by using RT-PCR. The sequence analysis suggested that the chromophore, consensus amino acids, and secondary structure of 11 β-strands of hmGFP were similar to those of GFP from other species. The recombinant hmGFP protein with high purity was obtained by the fusion expression of pETTRX-hmGFP in Escherichia coli and subsequent purification. The pH sensitivity and fluorescence spectroscopy of recombinant hmGFP were characterized. The excitation spectrum of recombinant hmGFP has a rather wide major peak with a maximum at 490nm and a shoulder at 420nm, and its emission spectrum at 510nm. The expression of hmGFP and the chimera IPL∼hmGFP in CHO cells has shown that the fusion protein IPL∼hmGFP has retained the normal membrane targeting of the IPL from Dasyatis akajei, as well as maintaining fluorescent properties similar to those of native hmGFP, suggesting a promising prospect of the application in biotechnology research for the new protein.

Original languageEnglish (US)
Pages (from-to)879-885
Number of pages7
JournalBiochemical and biophysical research communications
Volume301
Issue number4
DOIs
StatePublished - Feb 21 2003
Externally publishedYes

Keywords

  • Eukaryotic expression
  • Functional cloning
  • Fusion protein
  • GFP homolog
  • Prokaryotic expression

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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