Cloning, expression and purification of Microcystis viridis lectin in Escherichia coli.
Microcystis viridis lectin (MVL), a sugar-binding protein originally isolated from freshwater blue-green algae Microcystis viridis, has been reported to have potent anti-HIV activity. In this paper, we described the expression and purification of recombinant-MVL (R-MVL) gene in E. coli. The results demonstrated that the R-MVL in shake flask cultures was primarily expressed either in the form of inclusion bodies at 37°C or in the soluble fraction at 23 °C. Secondly, a one-step purification based on nickel-affinity chromatography was employed and 15 mg of highly purified (>95%) R-MVL from 1 l of cell cultures was yielded. The purified R-MVL was then subjected to MALDI-TOF-MS analysis for protein identification. In conclusion, for the first time, the R-MVL was successfully cloned and expressed in E. coli, which is useful for further study and large-scale cost-effective production of MVL protein.
Li
Yuqin
Y
College of Light Industry and Food Sciences, South China University of Technology, 381 Wushan Road, Guangzhou 510640, China.
Liao
Xiaoxia
X
Chen
Gu
G
Yap
Yeeleng
Y
Zhang
Xuewu
X
D001483:Base Sequence; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000235:genetics; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D037102:Lectins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D046931:Microcystis / Q000235:genetics*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification
D004274:DNA, Recombinant; D037102:Lectins; D011994:Recombinant Proteins