ISOLATION AND OVEREXPRESSION IN ESCHERICHIA-COLI OF THE FLAVODOXIN GENE FROM ANABAENA-PCC-7119

被引:79
作者
FILLAT, MF [1 ]
BORRIAS, WE [1 ]
WEISBEEK, PJ [1 ]
机构
[1] UNIV UTRECHT, DEPT MOLEC CELL BIOL, 3584 CH UTRECHT, NETHERLANDS
关键词
D O I
10.1042/bj2800187
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The gene coding for flavodoxin from Anabaena PCC 7119 was cloned by using the polymerase chain reaction (PCR). The gene is transcribed into a 1250-base transcript. The expression of the flavodoxin gene was analysed and found to be regulated at the transcriptional level by the availability of iron. The PCR-amplified gene was cloned into the expression vector pTrc 99b and expressed in Escherichia coli. High concentrations of flavodoxin were found (20% of total protein). The recombinant protein was purified from the cytosolic fraction of the cells and it exhibited properties identical with those of the wild-type Anabaena flavodoxin.
引用
收藏
页码:187 / 191
页数:5
相关论文
共 35 条
  • [21] Maniatis T., 1982, MOL CLONING
  • [22] IDENTIFICATION OF BLUE-GREEN-ALGAL NITROGEN-FIXATION GENES BY USING HETEROLOGOUS DNA HYBRIDIZATION PROBES
    MAZUR, BJ
    RICE, D
    HASELKORN, R
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1980, 77 (01): : 186 - 190
  • [23] HYBRIDIZATION OF NUCLEIC-ACIDS IMMOBILIZED ON SOLID SUPPORTS
    MEINKOTH, J
    WAHL, G
    [J]. ANALYTICAL BIOCHEMISTRY, 1984, 138 (02) : 267 - 284
  • [24] MULLIS KB, 1987, METHOD ENZYMOL, V155, P335
  • [25] RICE D, 1982, J BIOL CHEM, V257, P3157
  • [26] RIPPKA R, 1988, METHOD ENZYMOL, V167, P28
  • [27] PRIMER-DIRECTED ENZYMATIC AMPLIFICATION OF DNA WITH A THERMOSTABLE DNA-POLYMERASE
    SAIKI, RK
    GELFAND, DH
    STOFFEL, S
    SCHARF, SJ
    HIGUCHI, R
    HORN, GT
    MULLIS, KB
    ERLICH, HA
    [J]. SCIENCE, 1988, 239 (4839) : 487 - 491
  • [28] CONSEQUENCES OF THE IRON-DEPENDENT FORMATION OF FERREDOXIN AND FLAVODOXIN ON PHOTOSYNTHESIS AND NITROGEN-FIXATION ON ANABAENA STRAINS
    SANDMANN, G
    PELEATO, ML
    FILLAT, MF
    LAZARO, MC
    GOMEZMORENO, C
    [J]. PHOTOSYNTHESIS RESEARCH, 1990, 26 (02) : 119 - 125
  • [29] DNA SEQUENCING WITH CHAIN-TERMINATING INHIBITORS
    SANGER, F
    NICKLEN, S
    COULSON, AR
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1977, 74 (12) : 5463 - 5467
  • [30] SHAH VK, 1988, ARCH MICROBIOL, V258, P12064