High-Level Expression in Escherichia coli and Purification of the Membrane-Bound Form of Cytochrome b5

2000 ◽  
Vol 19 (1) ◽  
pp. 173-178 ◽  
Author(s):  
Scott B. Mulrooney ◽  
Lucy Waskell
Keyword(s):  
Escherichia Coli ◽  
Cytochrome B5 ◽  
High Level Expression ◽  
Expression In Escherichia Coli ◽  
Membrane Bound ◽  
High Level

High-level expression in Escherichia coli of calcium-binding domains of an embryonic sea urchin protein

Gene ◽  
1984 ◽  
Vol 31 (1-3) ◽  
pp. 155-164 ◽  
Author(s):  
Mark Muesing ◽  
Clifford D. Carpenter ◽  
William H. Klein ◽  
Barry Polisky
Keyword(s):  
Escherichia Coli ◽  
Sea Urchin ◽  
Calcium Binding ◽  
High Level Expression ◽  
Expression In Escherichia Coli ◽  
Binding Domains ◽  
High Level

Recombinant mouse prolactin: expression in Escherichia coli, purification and biological activity

1992 ◽  
Vol 8 (2) ◽  
pp. 165-172 ◽  
Author(s):  
M. Yamamoto ◽  
T. Harigaya ◽  
T. Ichikawa ◽  
K. Hoshino ◽  
K. Nakashima
Keyword(s):  
Escherichia Coli ◽  
Biological Activity ◽  
Amino Acid ◽  
Multicopy Plasmid ◽  
High Level Expression ◽  
E Coli ◽  
Expression In Escherichia Coli ◽  
Oligonucleotide Duplex ◽  
Efficient Expression ◽  
High Level

ABSTRACT Transformation of Escherichia coli cells with a recombinant plasmid containing modified mouse prolactin (mPRL) cDNA and a pKK223-3 vector resulted in efficient expression of mPRL protein. Cloned mPRL cDNA was modified by removing the 5′ non-translating sequence as well as the sequence which encoded the signal peptide of preprolactin for recombination. In addition, approximately 100 nucleotides of the 5′-terminal region of the cDNA, which include the ATG initiation codon and the following 31 codons of mature mPRL, were replaced by a chemically synthesized oligonucleotide duplex. The sequence of this duplex was chosen to be rich in AT without changing the amino acid sequence of the protein. The modified cDNA was finally inserted into the multicopy plasmid, pUC19, before high-level expression of mPRL in E. coli cells was obtained. Western blotting analysis of total protein from transformed E. coli cells showed that both 23 and 16kDa peptides were recognized by specific mPRL antisera. The purified and refolded 23 kDa protein exhibited a growth-stimulating effect on rat Nb 2 Node lymphoma cells, and was very similar to that of natural pituitary PRL.

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