Here is a standard blurb regarding the MET3 plasmid our lab uses for
expression of heterologous genes.
The MET3 gene was obtained from Harry Mountain at the University of
Sheffield in England. The plasmid contains a Bgl II to Eco RI
fragment of the MET3 gene ligated into pUC19 cut with Bam HI and Eco
RI. This contains the entire promoter of MET3 and a fragment of the
5' end of the MET3 coding region. Dr. Mountain has suggested using
an Eco RV site that lies at -68 relative to the AUG for fusion of this
promoter to heterologous genes. He has successfully used this for the
expression of the cytotoxin gene from the MET3 promoter and the gene
showed the proper methionine regulation. Other constructs for
heterologous expression used the Eco RI site to express MET3 chimaeric
proteins. This worked well for LacZ and URA3.
The voice phone number for Harry Mountain is 011 44 0742 768555 ext 6186
The FAX number is 011 44 0742 728697.
Here are the MET3 gene references:
Cherest et al. Gene, vol 34 (1985) 279-281.
Cherest et al. Mol Gen Genet, vol 210 (1987) 307-313.
Korch et al. Mol Gen Genet, vol 229 (1991) 96-108.
Mountain et al. Yeast, vol 7 (1991) 781-803.
Mountain and Korch Yeast, vol 7 (1991) 873-880.
ER | Bob Litt (rjl4 at midway.uchicago.edu)
\/ | Segev "if it's secreted, we like it" Lab
Golgi | Univ of Chicago Dept of Molec Genetics and Cell Biol