Expression and purification of Src I from sea anemone Sagartia rosea as a recombinant non-fusion protein

Xiaoyu Jiang, Lisheng Peng, Wenli Yang, Xiaojiang Tang, Wei Liu, Anlong Xu

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

The cDNA coding for an acidic actinoporin, Sagartia rosea cytolysin I (Src I), has been isolated, cloned into pGEM-T Easy Vector, and sequenced. The region encoding matured Src I was also cloned into prokaryotic expression vector pBV220 and expressed in Escherichia coli as a non-fusion protein in the form of inclusion body. Through washing and denaturation-renaturation step, the recombinant Src I was purified by Q Sepharose Fast Flow ion exchange chromatography and Phenyl Sepharose hydrophobic interaction chromatography. The two-step purification of Src I was effective, simple, and suitable for isolating large amount of high purity (above 95%) recombinant Src I.

Original languageEnglish (US)
Pages (from-to)161-166
Number of pages6
JournalProtein Expression and Purification
Volume32
Issue number1
DOIs
StatePublished - Nov 2003
Externally publishedYes

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Sea Anemones
Recombinant Fusion Proteins
Perforin
Purification
Chromatography
Proteins
Denaturation
Inclusion Bodies
Ion Exchange Chromatography
Hydrophobic and Hydrophilic Interactions
Washing
Sepharose
Escherichia coli
Ion exchange
Complementary DNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

Expression and purification of Src I from sea anemone Sagartia rosea as a recombinant non-fusion protein. / Jiang, Xiaoyu; Peng, Lisheng; Yang, Wenli; Tang, Xiaojiang; Liu, Wei; Xu, Anlong.

In: Protein Expression and Purification, Vol. 32, No. 1, 11.2003, p. 161-166.

Research output: Contribution to journalArticle

Jiang, Xiaoyu ; Peng, Lisheng ; Yang, Wenli ; Tang, Xiaojiang ; Liu, Wei ; Xu, Anlong. / Expression and purification of Src I from sea anemone Sagartia rosea as a recombinant non-fusion protein. In: Protein Expression and Purification. 2003 ; Vol. 32, No. 1. pp. 161-166.
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