title: Systematic high-yield production of human secreted proteins in Escherichia coli.
abstract: Human secreted proteins play a very important role in signal transduction. In order to study all potential secreted proteins identified from the human genome sequence, systematic production of large amounts of biologically active secreted proteins is a prerequisite. We selected 25 novel genes as a trial case for establishing a reliable expression system to produce active human secreted proteins in Escherichia coli. Expression of proteins with or without signal peptides was examined and compared in E. coli strains. The results indicated that deletion of signal peptides, to a certain extent, can improve the expression of these proteins and their solubilities. More importantly, under expression conditions such as induction temperature, N-terminus fusion peptides need to be optimized in order to express adequate amounts of soluble proteins. These recombinant proteins were characterized as well-folded proteins. This system enables us to rapidly obtain soluble and highly purified human secreted proteins for further functional studies.
authors.affiliation: National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Xueyu
authors.initials: X
authors.lastname: Dai
authors.forename: Qiang
authors.initials: Q
authors.lastname: Chen
authors.forename: Min
authors.initials: M
authors.lastname: Lian
authors.forename: Yanfeng
authors.initials: Y
authors.lastname: Zhou
authors.forename: Mo
authors.initials: M
authors.lastname: Zhou
authors.forename: Shanyun
authors.initials: S
authors.lastname: Lu
authors.forename: Yunjia
authors.initials: Y
authors.lastname: Chen
authors.forename: Jingchu
authors.initials: J
authors.lastname: Luo
authors.forename: Xiaocheng
authors.initials: X
authors.lastname: Gu
authors.forename: Ying
authors.initials: Y
authors.lastname: Jiang
authors.forename: Ming
authors.initials: M
authors.lastname: Luo
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Zheng
mesh_terms: D002874:Chromosome Mapping / Q000379:methods*; D003001:Cloning, Molecular / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000096:biosynthesis* / Q000235:genetics; D024861:Genetic Enhancement / Q000379:methods*; D006801:Humans; D015202:Protein Engineering / Q000379:methods*; D021382:Protein Sorting Signals / Q000235:genetics*
chemical_list: D029968:Escherichia coli Proteins; D021382:Protein Sorting Signals