Product Details

Purity

Greater than 90% as determined by SDS-PAGE.

Target Names

Uniprot No.

Q38087

Research Area

Others

Alternative Names

(Gp43)

Species

Escherichia phage RB69 (Bacteriophage RB69)

Source

E.coli

Expression Region

103-340aa

Target Protein Sequence

YDHTKIRVANFDIEVTSPDGFPEPSQAKHPIDAITHYDSIDDRFYVFDLLNSPYGNVEEWSIEIAAKLQEQGGDEVPSEIIDKIIYMPFDNEKELLMEYLNFWQQKTPVILTGWNVESFDIPYVYNRIKNIFGESTAKRLSPHRKTRVKVIENMYGSREIITLFGISVLDYIDLYKKFSFTNQPSYSLDYISEFELNVGKLKYDGPISKLRESNHQRYISYNIIDVYRVLQIDAKRQF
Note: The complete sequence may include tag sequence, target protein sequence, linker sequence and extra sequence that is translated with the protein sequence for the purpose(s) of secretion, stability, solubility, etc.
If the exact amino acid sequence of this recombinant protein is critical to your application, please explicitly request the full and complete sequence of this protein before ordering.

Mol. Weight

35.4 kDa

Protein Length

Partial

Tag Info

N-terminal 10xHis-tagged and C-terminal Myc-tagged

Form

Liquid or Lyophilized powder
Note: We will preferentially ship the format that we have in stock, however, if you have any special requirement for the format, please remark your requirement when placing the order, we will prepare according to your demand.

Buffer

If the delivery form is liquid, the default storage buffer is Tris/PBS-based buffer, 5%-50% glycerol. If the delivery form is lyophilized powder, the buffer before lyophilization is Tris/PBS-based buffer, 6% Trehalose.

Reconstitution

We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Please reconstitute protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL.We recommend to add 5-50% of glycerol (final concentration) and aliquot for long-term storage at -20°C/-80°C. Our default final concentration of glycerol is 50%. Customers could use it as reference.

Troubleshooting and FAQs

Protein FAQs

Storage Condition

Store at -20°C/-80°C upon receipt, aliquoting is necessary for mutiple use. Avoid repeated freeze-thaw cycles.

Shelf Life

The shelf life is related to many factors, storage state, buffer ingredients, storage temperature and the stability of the protein itself.
Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. The shelf life of lyophilized form is 12 months at -20°C/-80°C.

Lead Time

3-7 business days

Notes

Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.

Datasheet & COA

Please contact us to get it.

Description

The expression of recombinant Escherichia phage RB69 (Bacteriophage RB69) DNA-directed DNA polymerase (Gp43) protein involves transfecting e.coli cells with a DNA expression vector carrying the gene for the protein of interest (103-340aa). After transfection, the cells are cultured to produce the desired protein. A N-terminal 10xHis tag and C-terminal Myc tag is fused to the protein. The recombinant Escherichia phage RB69 (Bacteriophage RB69) DNA-directed DNA polymerase (Gp43) protein is collected and purified from the cell lysate through affinity purification, attaining a purity greater than 90%, as determined by SDS-PAGE.

Target Background

Function

Replicates the viral genomic DNA. This polymerase possesses two enzymatic activities: DNA synthesis (polymerase) and an exonucleolytic activity that degrades single-stranded DNA in the 3'- to 5'-direction for proofreading purpose.

Gene References into Functions

Co(2+) and Mn(2+) enhanced ground-state binding of both correct and incorrect dNTPs to RB69pol:dideoxy-terminated primer-template complexes. PMID: 27096230
Data indicate that alanine replacement of residues distant from the active site of the replicative RB69 DNA polymerase renders the enzyme incapable of sustaining phage replication in vivo. PMID: 24116139
structures of the various RB69pol ternary complexes can be used to rationalize the results obtained from pre-steady-state kinetic assays[review] PMID: 24720884
Determined is the crystal structure of the dATP/tC(o)-containing ternary complex of the RB69 DNA polymerase Y567A mutant at 1.9 A resolution; the incoming dATP formed two hydrogen bonds with an imino-tautomerized form of tC(o). PMID: 22616982
evaluated the contribution of minor groove hydrogen bonding interactions with RB69pol PMID: 22571765
The miscoding potential of 5-hydroxycytosine arises due to template instability in the DNA polymerase active site. PMID: 22026756
The authors show that the S565G/Y567A mutant generally had greater base selectivity than the Y567A mutant and that the kinetic parameters for dNMP insertion, excision of the 3'-terminal nucleotide residue, and primer extension beyond a mispair differed. PMID: 21216248
The structure of an RB69 pol ternary complex at 1.8 A is reported. A network of five highly ordered, buried water molecules can be seen to interact with the N3 and O2 atoms in the minor groove of the DNA duplex. PMID: 21158418
When Y567A and S565G replacements were combined, mutator activity was strongly decreased compared to that with Y567A replacement alone. PMID: 20950625
Data describe the crystal structures of the pol alpha family RB69 DNA polymerase with DNA containing the two most prevalent, spontaneously generated premutagenic lesions PMID: 15057282
Results describe the structure of the bacteriophage RB69 replicative DNA polymerase attempting to process an abasic site analog. PMID: 15057283
The Y619F substitution would disrupt the hydrogen bond network at the primer terminus and may affect the alignment of the 3' primer terminus at the polymerase active site, slowing chemistry and overall DNA synthesis. PMID: 17321543
Binding of a metal ion to the A site is required for the nucleotidyl transfer reaction, but is insufficient to initiate the enzyme isomerization. Binding of a dNTP, in the absence of a metal ion, is sufficient to induce this conformational change. PMID: 19228036
The effect of A and B metal ion site occupancy on the rates of Fingers closing and on the affinity of Rh.dTTP for the E.D binary complex of the catalytic site of RB69 DNA polymerase was studied. PMID: 19228037

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Protein Families

DNA polymerase type-B family

Database Links

KEGG: vg:1494172

Code	CSB-EP655714ELM
Abbreviation	Recombinant Enterobacteria phage RB69 DNA polymerase protein, partial
MSDS	MSDS Datasheet
Size	$388
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Image	(Tris-Glycine gel) Discontinuous SDS-PAGE (reduced) with 5% enrichment gel and 15% separation gel.
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Recombinant Enterobacteria phage RB69 DNA polymerase (43), partial

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