Recombinant mouse TDG

Catalog # : EPX-003-RBV

Source : Mouse

Expressed in : SF9 cells

Quantity : 10µg of enzymatically active recombinant TDG at 0.5µg /µl

Background:

TDG is a G/T mismatch-specific thymine-DNA glycosylase (1, 2). In the DNA of higher eukaryotes, hydrolytic deamination of 5-methylcytosine to thymine leads to the formation of G/T mismatches. This enzyme corrects G/T mispairs to G/C pairs (3). It is capable of hydrolyzing the carbon-nitrogen bond between the sugar-phosphate backbone of the DNA and a mispaired thymine (3). In addition to the G/T, it can remove thymine also from C/T and T/T mispairs in the order G/T >> C/T > T/T. It has no detectable activity on apyrimidinic sites and does not catalyze the removal of thymine from A/T pairs or from single-stranded DNA. It can also remove uracil and 5-bromouracil from mispairs with guanine.

Protein details:

Recombinant mouse N-terminal His tagged TDG was produced in SF9 insect cells, purified using FPLC and formulated in a storage buffer containing 20mM Tris-Cl pH 7.6, 1mM EDTA, 0.15 M NaCl, 10% glycerol, 0.5mM PMSF and 1mM DTT. Protein concentration was determined by spectrometry. >95% purity by SDS-PAGE.

Quality control:

In vitro glycosylase assay:

A  70 bp DNA containing a G/T mismatch was incubated or not with recombinant MBD4 for 30 minutes at 37°C. The reaction products were treated with NaOH and run on PAGE under denaturing conditions. Note that a 35 bp cut product was generated in the presence of MBD4.

Storage:

-80°C

Guarantee:

For research use only. Products guaranteed stable for 2 years from date of receipt when stored properly.

Purity:

>98% purity by SDS PAGE.

Sequence:

MGSSHHHHHHSSGLVPRGSHMLEPGGMDAEAARSYSLEQVQALYSFPFQQ
MMAEVPNMAVTTGQQVPAVAPNMATVTEQQVPADAPVQEPAPEAPKRRKR
KPRAAEPQEPVEPKKPATSKKSGKSTKSKEKQEKITDAFKVKRKVDRFNG
VSEAELLTKTLPDILTFNLDIVIIGINPGLMAAYKGHHYPGPGNHFWKCL
FMSGLSEVQLNHMDDHTLPGKYGIGFTNMVERTTPGSKDLSSKEFREGGR
ILVQKLQKYQPRIAVFNGKCIYEIFSKEVFGVKVKNLEFGLQPHKIPDTE
TLCYVMPSSSARCAQFPRAQDKVHYYIKLKDLRDQLKGIERNTDVQEVQY
TFDLQLAQEDAKKMAVKEEKYDPGYEAAYGGAYGENPCNGEPCGIASNGL
TAHSAEPRGEATPGDVPNGQWMAQSFAEQIPSFNNCGTREQEEESHA*

References:

1.    Neddermann and Jiricny. J. Biol. Chem. 268:21218-21224(1993)
2.    Neddermann et al., J. Biol. Chem. 271:12767-12774(1996)
3.    Neddermann and Jiricny. Proc. Natl. Acad. Sci. U.S.A. 91:1642-1646(1994)