Unit of Genetics and Molecular Biology, Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia.
Genet Mol Biol. 2011 Jul;34(3):464-70. doi: 10.1590/S1415-47572011005000022. Epub 2011 Jul 1.
The Pichia pastoris expression system was used to produce recombinant human erythropoietin, a protein synthesized by the adult kidney and responsible for the regulation of red blood cell production. The entire recombinant human erythropoietin (rhEPO) gene was constructed using the Splicing by Overlap Extension by PCR (SOE-PCR) technique, cloned and expressed through the secretory pathway of the Pichia expression system. Recombinant erythropoietin was successfully expressed in P. pastoris. The estimated molecular mass of the expressed protein ranged from 32 kDa to 75 kDa, with the variation in size being attributed to the presence of rhEPO glycosylation analogs. A crude functional analysis of the soluble proteins showed that all of the forms were active in vivo.
利用毕赤酵母表达系统生产重组人促红细胞生成素，这种蛋白由成人肾脏合成，负责调节红细胞的生成。通过 PCR 重叠延伸拼接（SOE-PCR）技术构建整个重组人促红细胞生成素（rhEPO）基因，通过毕赤酵母表达系统的分泌途径进行克隆和表达。重组促红细胞生成素在毕赤酵母中成功表达。表达蛋白的估计分子质量范围为 32kDa 至 75kDa，大小的变化归因于 rhEPO 糖基化类似物的存在。对可溶性蛋白的粗功能分析表明，所有形式的蛋白在体内均具有活性。
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