23 Jul Yeast expression carrier for combined bone sialic acid protein, purification method and its application
Abstract
The invention discloses an expressing carrier of fuse bone sialic acid protein yeast, purified method and appliance in gene engineering and microbe fermentation engineering technical domain, which comprises the following steps: designing gene special primer according to human bone sialic acid protein (hbsp) gene sequence; augmenting cDNA of BSP from total RNA of human femur exine tissue with RT-PCR technique; cloning to pBluescript II RI plasmid; constructing retooling plasmid pB-hbsp; designing PCR primer according to pichia secretary type expressing plasmid pPICZaA and hbsp gene sequence; choosing retooling plasmid pB-hbsp as mold; proceeding PCR augment; restructuring PCR product and plasmid pPICZaA; constructing plasmid pPICZaA-hbsp of secretary type fuse expressing rhBSP-His6; purifying the fuse expressing rhBSP-His6 with fixed metallic ion affinity chromatography (IMAC); reaching ionophresis pure of goal protein rhBSP. This invention provides a new approach for acquisition of BSP, which lays the foundation for further studying BSP immunology.
Year: 2007
Country: CN
Doc No: 101063146