[Fusion expression of human thymosin alpha 1 in Escherichia coli].
Engineering E. coli strain, BL21 (DE3)/pGEX-4T-human Thymosin alpha 1, was constructed by oligonucleotide annealing and PCR amplifying the target gene, then ligating it with pGEX-4T-3 vector and transferring into BL21 host. The yield of fusion protein of GST-Thymosin alpha 1 expressed from BL21 (DE3)/pGEX-4T-thymosin alpha 1 is about 35%-40% of total protein after fermentation. Following the simple cut of thrombin or CNBr, about 0.2 g/L thymosin alpha 1 can be harvested. The product is checked by MS and activity test, which indicates that the recombinant product has full biological activity of native thymosin alpha 1.
Xiu
Zhao-Yang
ZY
Research Center of Biotechnology, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences, Shanghai 200233, China.
Yu
Ying
Y
Chen
Chang-Qing
CQ
D004926:Escherichia coli / Q000235:genetics*; D005818:Genetic Engineering; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D000077596:Thymalfasin; D013947:Thymosin / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification
D011993:Recombinant Fusion Proteins; D013947:Thymosin; D000077596:Thymalfasin