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.
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页码:187 / 191
页数:5
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