| 產(chǎn)品名稱 |
pMT100/pSU77 [NEB548] |
| 商品貨號(hào) |
B229535 |
| Designations |
pMT100/pSU77 [NEB548] |
| GenBank Number |
X17195
|
| Species |
Escherichia coli (Migula) Castellani and Chalmers |
| Depositors |
New England Biolabs, Inc., I Schildkraut, New England Biolabs, Inc. |
| Applications |
produces protein modification methylase SssI |
| Vector |
pMT100 construct size (kb): 4.5
pSU77 construct size (kb): 4.6 |
| Insert |
DNA: genomic hsdM SssI insert lengths(kb): 1.538 Gene product: tRNA Trp; trpT176, su7 opal suppressor (UGA) [hsdM] Target Gene: modification methylase SssI supU gene insert length (kb): 0.264 |
| Insert Size (kb) |
1.538 and 0.264 |
| Media |
ATCC Medium 1065 (see below) plus ampicillin (50 mcg/ml) plus chloramphenicol (20 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. |
| Shipping Information |
Shipped: ?Freeze dried E. coli containing the plasmids |
| Comments |
There are 2 plasmids in this E. coli, pSU77 and pMT100. The pMT100 phagemid contains the modification methylase SssI from Spiroplasma sp. The pSU77 construct is necessary to suppress UGA codons at positions 229, 349, 595 and 709 that function to terminate translation in E. coli. The insert of plasmid pMT100 contains the following restriction sites (approximate kb from the 5' end): EcoRI--1.10, 1.40; EcoRV--0.81; HindIII--0.15, 0.97; SacI--0.67; XbaI--0.30. Expression of the suppressor tRNA is regulated by a lacUV5 promoter and a rrnC terminator. Restriction digests of pSU77 give the following sizes (kb): KpnI/NotI--4.6; PstI/XbaI--4.6; PstI--4.6; HindIII--4.6; EcoRI--4.2, 0.3. Restriction digests pMT100 give the following sizes (kb): KpnI/NotI--3.0, 1.5; PstI/XbaI--3.0, 1.2, 0.4; PstI--4.6; HindIII--3.0, 0.9, 0.7; EcoRI--2.3, 2.0, 0.17. |
| Classification |
Enterobacteriaceae, Escherichia |
| References |
Razin A, et al. DNA encoding Spiroplasma sp. DNA methylase. US Patent 5,296,371 dated Mar 22 1994
Raftery LA, et al. Defined set of cloned termination suppressors: in vivo activity of isogenetic UAG, UAA, and UGA suppressor tRNAs. J. Bacteriol. 158: 849-859, 1984. PubMed: 6327642
Renbaum P, et al. Cloning, characterization, and expression in Escherichia coli of the gene coding for the CpG DNA methylase from Spiroplasma sp. strain MQ1(M.SssI). Nucleic Acids Res. 18: 1145-1152, 1990. PubMed: 2181400
|
| 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. |
