抗癌基因Rb在大肠杆菌中的表达

EXPRESSION OF RETINOBLASTOMA SUSCEPTIBILITY GENE IN E.COLI

Ｒｂ基因ｃＤＮＡ８４４ｂｐ片段，与高效原核表达载体ｐＢＶ２２１重组，构建了重组表达质粒ｐＲＢ８４４，在大肠杆菌中成功地表达了Ｒｂ基因的产物，其分子量约３２ｋＤ，表达量约占菌体总蛋白的１０％－１５％，初步纯化其纯度可达５０％－６０％，进一步纯化其纯度可达９８％以上。经家兔免疫后，获得了抗Ｒｂ基因产物的抗体，这为Ｒｂ基因的进一步深入研究提供了条件。

b cDNA 844bp fragment was inserted into the expression vector pBV221. The recombinant plasmid pRB844 highly expressed a 32kD protein in E.coli BL21. The percentage is 10%-15% of total bacterial protein. Primary purity of the expressed protein through ureapurification is 50%-60%,the puriry could reach more than 98% through further purification.Polyclonal rabbit serum against the Rb protein is also prepared.