EXPRESSION OF SACCHAROMYCES-CEREVISIAE INORGANIC PYROPHOSPHATASE IN ESCHERICHIA-COLI

被引:0
|
作者
KURILOVA, SA
VOROBJEVA, NN
NAZAROVA, TI
AVAEVA, SM
机构
[1] MOSCOW MV LOMONOSOV STATE UNIV,AN BELOZERSKY INST PHYSICOCHEM BIOL,MOSCOW 119899,RUSSIA
[2] MOSCOW MV LOMONOSOV STATE UNIV,DEPT CHEM,MOSCOW 119899,RUSSIA
关键词
SACCHAROMYCES-CEREVISIAE; INORGANIC PYROPHOSPHATASE; GENE; RECOMBINANT VECTOR; EXPRESSION; ESCHERICHIA-COLI; PCR;
D O I
10.1016/0014-5793(93)80670-P
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
A segment of DNA encoding Saccharomyces cerevisiae inorganic pyrophosphatase (ppa gene) was amplified by the polymerase chain reaction. The pSCH1 and pSCB6 plasmids containing the ppa gene were obtained. Transformation of the E coli BL21 strain with the resulting recombinant plasmids and selection of clones having extremely high expression of inorganic pyrophosphatase (PPase) were carried out. Superproduction of recombinant S. cerevisiae PPase up to 50% of the total bacterial protein was achieved. The enzyme was readily obtained and purified to homogeneity with the use of a simple purification technique. This work is the first description of S. cerevisiae PPase superproducer creation.
引用
收藏
页码:280 / 282
页数:3
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