標題: Expression and purification of human placenta lactogen in Escherichia coli
作者: Lan, PC
Tseng, CF
Lin, MC
Chang, CA
生物科技學系
Department of Biological Science and Technology
關鍵字: human placental lactogen;molecular cloning;protein expression
公開日期: 1-四月-2006
摘要: There are many growth factors secreted by placenta including growth hormone, placenta lactogen (PL), prolactin, follicle stimulating hormone, luteinizing hormone, thyroid stimulating hormone, and chorionic gonadotropin. For a systematic study of how these growth factors work together to result in the various biological functions and future clinical applications, it is needed to produce enough quantities of each protein. In this paper, we report the cloning of human PL (hPL) and expression by Escherichia coli (E coli). Four kinds of expression vectors containing the hPL gene were transformed into several kinds of suitable host strains and grown at 37 and/or 30 degrees C. Determination of the yield of recombinant hPL by SDS-PAGE reveals that among the various conditions, pQE30-PL in E. coli strain M15[pREP4] expressed the largest amount of recombinant hPL at 37 degrees C. However, the expressed recombinant hPL was accumulated in inclusion body forms. The inclusion bodies were solubilized in 8 M urea and purified by a His(6) tagged affinity column under denaturing condition and the final yield of hPL was determined to be 48 mg/L. Intra-chain disulfide bonds could be formed either by oxidation in the refolding buffer or by air oxidation in the presence of urea. The biological activity was examined by the fact that hPL could stimulate erythroid maturation by the formation of hemoglobin in K-562 cells in the presence of erythropoietin. Initial optimization studies resulted in the production of 282.4 mg/L of hPL. (c) 2005 Elsevier Inc. All rights reserved.
URI: http://dx.doi.org/10.1016/j.pep.2005.08.019
http://hdl.handle.net/11536/12407
ISSN: 1046-5928
DOI: 10.1016/j.pep.2005.08.019
期刊: PROTEIN EXPRESSION AND PURIFICATION
Volume: 46
Issue: 2
起始頁: 285
結束頁: 293
顯示於類別:期刊論文


文件中的檔案:

  1. 000236828100014.pdf