Expression and purification of Src I from sea anemone Sagartia rosea as a recombinant non-fusion protein.
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.
Jiang
Xiaoyu
X
The Open Laboratory for Marine Functional Genomics of State High-Tech Development, Department of Biochemistry, College of Life Sciences, Sun Yat-sen (Zhongshan) University, Guangzhou 510275, People's Republic of China.
Peng
Lisheng
L
Yang
Wenli
W
Tang
Xiaojiang
X
Liu
Wei
W
Xu
Anlong
A
D000818:Animals; D002853:Chromatography, Liquid; D002942:Circular Dichroism; D003001:Cloning, Molecular; D003603:Cytotoxins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification* / Q000494:pharmacology; D004912:Erythrocytes / Q000166:cytology / Q000187:drug effects; D005822:Genetic Vectors / Q000235:genetics; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification* / Q000494:pharmacology; D012615:Sea Anemones / Q000737:chemistry / Q000235:genetics
D003603:Cytotoxins; D011994:Recombinant Proteins