PURIFICATION AND CHARACTERIZATION OF DIHYDROFOLATE-REDUCTASE OF PLASMODIUM-FALCIPARUM EXPRESSED BY A SYNTHETIC GENE IN ESCHERICHIA-COLI

被引：32

作者：

SANO, G ^{[1
]}

MORIMATSU, K ^{[1
]}

HORII, T ^{[1
]}

机构：

[1] OSAKA UNIV,DEPT PROTOZOOL & PARASITOL,MICROBIAL DIS RES INST,SUITA,OSAKA 565,JAPAN

来源：

MOLECULAR AND BIOCHEMICAL PARASITOLOGY | 1994年 / 63卷 / 02期

关键词：

DIHYDROFOLATE REDUCTASE; PLASMODIUM FALCIPARUM; SYNTHETIC GENE; DRUG DESIGN;

D O I：

10.1016/0166-6851(94)90062-0

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：

We have expressed the dihydrofolate reductase (DHFR) part of the DHFR-thymidylate synthetase complex of P. falciparum in Escherichia coli, by constructing a gene with synthetic oligonucleotides that changed the gene's codon usages. The induced expression in an E. coli cell of the synthetic gene yielded a product that constituted about 30% of the total bacterial protein. The product was precipitated in an inclusion body in a cell. Its enzymatic activity was restored after denaturation and renaturation procedures with guanidine-HCl. Recombinant DHFRs with Ser or Thr at position 108 were prepared. Kinetic characterization showed that the DHFR(Ser108) has less of an affinity for NADPH and dihydrofolate than the DHFR(Thr108)

引用

页码：265 / 273

页数：9

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