| Since interferon was discovered in 1957, it has been hailed as a significant antiviral agent. In the 1970s, interferon made a big splash as a symbol of recombinant gene technology and the medical breakthroughs it would bring. The natural antiviral and antitumor activity of interferon alpha and beta promoted investigation for therapeutic use. Interferon therapy has been successfully implemented for skin malignancies, viral infections, kaposi sarcoma, multiple sclerosis, leukima, et al.According to the relation between gene expression levels and synonymcodon usage, base composition in coding region,base pair correlation in a codon and base pair correlation between codons. Human Interferonα-2b gene was designed by using degenracy of codon. The designed gene was expected to be expressed highly in Pichia pastroris. Oligo DNA was synthesized to form complete human interferonα-2b gene fragment that was inserted into the vector pPIC9K. DNA sequencing show that Human interferonα-2b gene sequence of synthesis and insertion is completely consistent with that of the designed gene. Meantime, the expression plamid pPIC9K-IFNα-2b was constructed. By PCR analysis, construction was proved to be right. After electroporation of Pichia pastoris GS115, some high-copy transformants (His+Mut+) were picked up by Geneticin(G418). After 5 days induction, the result was tested by SDS-PAGE. SDS-PAGE result showed that IFNα-2b was expressed successfully.
|
PDF Full Text Request