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pMT100/pSU77 [NEB548]

货号 TS147381
中文名称 null
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产品名称: pMT100/pSU77 NEB548
商品货号: TS147381
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: xa0Freeze 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.