Construction of Integrative Expression Vector and Expression of Human Angiostatin in Hansenula polymorpha
汉森酵母表达载体的构建和人血管生成抑制素基因的表达

Hansenula polymorpha is a methylotrpphic yeast which can grow on methanol as the sole source of carbon and energy and is characterized by its easy manipulation in industry. The construction of an integrative expression vector and the expression of angiostatin in H.polymorpha have been described.The methanol oxidase(MOX) gene promoter and terminator of H.polymorpha was amplified by PCR and recombined with its leu gene and angiostatin gene on plasmid pSP72 resulting in the integrative vector pSMA17 which was then transformed into H.polymorpha A16(leu-).The positive transformant HpA16(pSMA17) was screened on SC medium without leucine.HpA16(pSMA17) was grown on YPGE medium to late log phase and induced with methanol.ELISA and SDS-PAGE results show that angiostatin was expressed and the expressed product secreted into the medium at 50mg/L fermentation broth(in the flasks).Western blot analysis Indicates that the recombinant angiostatin specifically reacts with antiserum against human plasminogen.