Induction conditions for maximizing recombinant staphylokinase expression in Escherichia coli

被引:4
作者
Yarzabal, A
Bastidas, M
Avilan, L
Cruz, J
Puig, J
机构
[1] Lab. de Biol. y Med. Experimental, Facultad de Ciencias, Universidad de Los Andes, Mérida
关键词
D O I
10.1023/A:1018330613369
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Staphylokinase, a profibrinolytic bacterial protein, was cloned into Escherichia coli, following the amplification of its gene via PCR. The amplificated gene was inserted in a pKK223-3 plasmid vector. The recombinant protein (STAR), expressed from a tac promoter, was obtained in the periplasmic space when IPTG was added to the culture medium. Both the concentration of the inducer as well as the growth phase of recombinant cells at which it was added affected the final yield of periplasmic STAR. The protein was purified by a one-step procedure in an acylated-plasminogen Sepharose coupled column.
引用
收藏
页码:633 / 637
页数:5
相关论文
共 18 条
[1]   CLONING AND EXPRESSION IN ESCHERICHIA-COLI, BACILLUS-SUBTILIS, AND STREPTOCOCCUS-SANGUIS OF A GENE FOR STAPHYLOKINASE - A BACTERIAL PLASMINOGEN-ACTIVATOR [J].
BEHNKE, D ;
GERLACH, D .
MOLECULAR & GENERAL GENETICS, 1987, 210 (03) :528-534
[2]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[3]   MECHANISMS OF ACTIVATION OF MAMMALIAN PLASMA FIBRINOLYTIC SYSTEMS WITH STREPTOKINASE AND WITH RECOMBINANT STAPHYLOKINASE [J].
COLLEN, D ;
VANHOEF, B ;
SCHLOTT, B ;
HARTMANN, M ;
GUHRS, KH ;
LIJNEN, HR .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1993, 216 (01) :307-314
[4]   ISOLATION AND CHARACTERIZATION OF NATURAL AND RECOMBINANT STAPHYLOKINASE [J].
COLLEN, D ;
SILENCE, K ;
DEMARSIN, E ;
DEMOL, M ;
LIJNEN, HR .
FIBRINOLYSIS, 1992, 6 (04) :203-213
[5]   PROLONGED INCUBATION IN CALCIUM-CHLORIDE IMPROVES THE COMPETENCE OF ESCHERICHIA-COLI-CELLS [J].
DAGERT, M ;
EHRLICH, SD .
GENE, 1979, 6 (01) :23-28
[6]   PLASMINOGEN - PURIFICATION FROM HUMAN PLASMA BY AFFINITY CHROMATOGRAPHY [J].
DEUTSCH, DG ;
MERTZ, ET .
SCIENCE, 1970, 170 (3962) :1095-+
[7]  
HAQQI T, 1992, NUC ACIDS RES, V20, P6247
[8]   CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].
LAEMMLI, UK .
NATURE, 1970, 227 (5259) :680-+
[9]  
LIJNEN HR, 1991, J BIOL CHEM, V266, P11826
[10]   ON THE MOLECULAR-INTERACTIONS BETWEEN PLASMINOGEN-STAPHYLOKINASE, ALPHA-2-ANTIPLASMIN AND FIBRIN [J].
LIJNEN, HR ;
VANHOEF, B ;
MATSUO, O ;
COLLEN, D .
BIOCHIMICA ET BIOPHYSICA ACTA, 1992, 1118 (02) :144-148