produces protein S-adenosyl-L-methionine hydrolase
Vector
Construct size (kb): 5.90
Insert
DNA: cDNA
Insert lengths(kb): 1.70
Gene product: S-adenosyl-L-methionine hydrolase
Target Gene: S-adenosyl-L-methionine hydrolase
Insert Size (kb)
1.700
Media
ATCC Medium 1065 (see below) plus kanamycin (5 mcg/ml) and chloramphenicol (5 mcg/ml)
ATCC Medium 1065:
Tryptone (Difco 0123), 10.0 g
Yeast Extract (Difco 0127), 5.0 g
NaCl, 10.0 g
Distilled water, 1.0 L
Biosafety Level
1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Comments
This is an expression construct of bacteriophage T3 S-adenosyl-L-methionine hydrolase (SAM) under the control of the Tn10 tet promoter in the pACYC184 vector.
Restriction endonuclease encoding cDNA clones are identified as those clones that grow well in the presence of chlortetracycline but grow poorly in its absence.
The host GUBCE-1 contains a dinD promoter operator/beta-galactosidase fusion that expresses beta-galactosidase in response to DNA damage.
The transformed host GUBCE-1/pGUBC2080 contains a p15A replicon based plasmid that is useful for recovering restriction endonuclease clones from cDNA libraries that have been constructed with compatible replicon (pMB1 based) vectors.
The SAM hydrolase gene product from GUBC-2080 is under tight regulation of the Tn10 tetR promoter. Transcription is easily controlled in a dose-dependent fashion by the inducer chlortetracycline.
References
Collier GB. Cloning restriction endonuclease genes by modulating methyltransferase activity. US Patent 5,451,519 dated Sep 19 1995
Disclosure
This material is cited in a US or other Patent and may not be used to
infringe the claims. Depending on the wishes of the Depositor, ATCC may be
required to inform the Patent Depositor of the party to which the material was
furnished. This material may not have been produced or characterized by ATCC.