HIGH-LEVEL SYNTHESIS IN ESCHERICHIA-COLI OF SHORTENED AND FULL-LENGTH HUMAN ACIDIC FIBROBLAST GROWTH-FACTOR AND PURIFICATION IN A FORM STABLE IN AQUEOUS-SOLUTIONS

被引:68
作者
ZAZO, M [1 ]
LOZANO, RM [1 ]
ORTEGA, S [1 ]
VARELA, J [1 ]
DIAZOREJAS, R [1 ]
RAMIREZ, JM [1 ]
GIMENEZGALLEGO, G [1 ]
机构
[1] CSIC,CTR INVEST BIOL,VELAZQUEZ 144,E-28006 MADRID,SPAIN
关键词
RECOMBINANT DNA; MESSENGER RNA SECONDARY STRUCTURES; POLYCISTRONIC MESSENGER TRANSLATION; INITIATOR METHIONINE PROCESSING; PROTEIN MODIFICATION BY REVERSED-PHASE CHROMATOGRAPHY;
D O I
10.1016/0378-1119(92)90400-J
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
A highly efficient expression for human acidic fibroblast growth factor (aFGF) has been assembled to direct the synthesis of both shortened and native full-length aFGF. The full-length aFGF-154 form of the protein had not been produced before in Escherichia coli by genetic engineering, and is obtained with its initiator methionine removed. The high production of the aFGF allows one to circumvent the use of reversed-phase chromatography (RPC) during the purification procedure. Here, it is shown that RPC, routinely used to obtain pure preparations of recombinant aFGF, modifies its chemical and physical properties in an unfavorable manner.
引用
收藏
页码:231 / 238
页数:8
相关论文
共 28 条
  • [11] ROLE OF CONFORMATIONAL-CHANGES IN THE ELUTION OF PROTEINS FROM REVERSED-PHASE HPLC COLUMNS
    KATZENSTEIN, GE
    VRONA, SA
    WECHSLER, RJ
    STEADMAN, BL
    LEWIS, RV
    MIDDAUGH, CR
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1986, 83 (12) : 4268 - 4272
  • [12] EXPRESSION OF SYNTHETIC GENES ENCODING BOVINE AND HUMAN BASIC FIBROBLAST GROWTH-FACTORS (BFGFS) IN ESCHERICHIA-COLI
    KNOERZER, W
    BINDER, HP
    SCHNEIDER, K
    GRUSS, P
    MCCARTHY, JEG
    RISAU, W
    [J]. GENE, 1989, 75 (01) : 21 - 30
  • [13] QUANTIFICATION OF CELLS CULTURED ON 96-WELL PLATES
    KUENG, W
    SILBER, E
    EPPENBERGER, U
    [J]. ANALYTICAL BIOCHEMISTRY, 1989, 182 (01) : 16 - 19
  • [14] Lakowicz, 1986, PRINCIPLES FLUORESCE
  • [15] Disulfide Bonds are Neither Required, Present, Nor Compatible with Full Activity of Human Recombinant Acidic Fibroblast Growth Factor
    Linemeyer, David L.
    Menke, John G.
    Kelly, Linda J.
    Disalvo, Jerry
    Soderman, Denis
    Schaeffer, M. -T.
    Ortega, Sagrario
    Gimenez-Gallego, Guillermo
    Thomas, Kenneth A.
    [J]. GROWTH FACTORS, 1990, 3 (04) : 287 - 298
  • [16] EXPRESSION IN ESCHERICHIA-COLI OF A CHEMICALLY SYNTHESIZED GENE FOR BIOLOGICALLY-ACTIVE BOVINE ACIDIC FIBROBLAST GROWTH-FACTOR
    LINEMEYER, DL
    KELLY, LJ
    MENKE, JG
    GIMENEZGALLEGO, G
    DISALVO, J
    THOMAS, KA
    [J]. BIO-TECHNOLOGY, 1987, 5 (09): : 960 - 965
  • [17] MASUI Y, 1983, EXPT MANIPULATION GE, P15
  • [18] OLIVER MH, 1989, J CELL SCI, V92, P513
  • [19] ORTEGA S, 1991, J BIOL CHEM, V266, P5842
  • [20] Sambrook J., 1989, MOL CLONING LAB MANU