Name: Recombinant Human DNA polymerase theta (POLQ), partial
Brand: CUSABIO
SKU: CSB-EP018321HU

Product Details

Purity

Greater than 85% as determined by SDS-PAGE.

Target Names

POLQ

Uniprot No.

O75417

Research Area

Epigenetics and Nuclear Signaling

Alternative Names

DNA polymerase eta

Species

Homo sapiens (Human)

Source

E.coli

Expression Region

321-554aa

Target Protein Sequence

GDEDHVVSLCYETICDNHSVLLFCPSKKWCEKLADIIAREFYNLHHQAEGLVKPSECPPVILEQKELLEVMDQLRRLPSGLDSVLQKTVPWGVAFHHAGLTFEERDIIEGAFRQGLIRVLAATSTLSSGVNLPARRVIIRTPIFGGRPLDILTYKQMVGRAGRKGVDTVGESILICKNSEKSKGIALLQGSLKPVRSCLQRREGEEVTGSMIRAILEIIVGGVASTSQDMHTYA
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

33.3 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

To produce recombinant Human POLQ protein, a well-established recombinant DNA technology is the key. A DNA template of POLQ was constructed with N-terminal 10xHis tag & C-terminal Myc tag using the technique. Once the template was made, the recombinant Human POLQ protein could be produced with it efficiently. CUSABIO has built a strict QC system to ensure quality. The expression region is 321-554aa of the Human POLQ. The purity of this recombinant is 85% determined by SDS-PAGE.

POLQ (POLH) is a protein coding gene that encodes DNA polymerase theta. According to some studies, POLQ may have the following features.
POLQ inhibits genomic instability and genomic rearrangements derived from DNA double-strand breaks. POLQ is a member of a similar cluster of up-regulated genes that are strongly associated with poor clinical outcomes in breast, ovarian and other types of cancer. Inhibition of POLQ is a compelling combination therapy for radiosensitization. POLQ-deficient mutants displayed hypersensitivity to H2O2-induced oxidative alkali damage, but not to UV light or cisplatin. Overexpression of POLQ leads to poor prognosis in patients with early breast cancer.

Target Background

Function

DNA polymerase that promotes microhomology-mediated end-joining (MMEJ), an alternative non-homologous end-joining (NHEJ) machinery triggered in response to double-strand breaks in DNA. MMEJ is an error-prone repair pathway that produces deletions of sequences from the strand being repaired and promotes genomic rearrangements, such as telomere fusions, some of them leading to cellular transformation. POLQ acts as an inhibitor of homology-recombination repair (HR) pathway by limiting RAD51 accumulation at resected ends. POLQ-mediated MMEJ may be required to promote the survival of cells with a compromised HR repair pathway, thereby preventing genomic havoc by resolving unrepaired lesions. The polymerase acts by binding directly the 2 ends of resected double-strand breaks, allowing microhomologous sequences in the overhangs to form base pairs. It then extends each strand from the base-paired region using the opposing overhang as a template. Requires partially resected DNA containing 2 to 6 base pairs of microhomology to perform MMEJ. The polymerase activity is highly promiscuous: unlike most polymerases, promotes extension of ssDNA and partial ssDNA (pssDNA) substrates. Also exhibits low-fidelity DNA synthesis, translesion synthesis and lyase activity, and it is implicated in interstrand-cross-link repair, base excision repair and DNA end-joining. Involved in somatic hypermutation of immunoglobulin genes, a process that requires the activity of DNA polymerases to ultimately introduce mutations at both A/T and C/G base pairs.

Gene References into Functions

Cells doubly deficient in Pol theta; and Lig4 exhibit 100% gene-targeting efficiency because of virtually no random integration events. PMID: 28695890
Data suggest that translesion DNA synthesis mediated by (1) POLI-dependent pathway (2) REV1- and POLN-dependent pathway, or (3) POLtheta-dependent pathway occur in predominantly error-free manner in human cells. (POLI = DNA polymerase iota; REV1 = DNA repair protein-REV1; POLN = DNA polymerase nu; POLtheta = DNA polymerase theta) PMID: 29330301
results suggest that bypass of Tg by Pol theta; results in mutations opposite the lesion, as well as frameshift mutations PMID: 29243925
This article summarizes work on the expression and purification of the full-length protein, and then focus on the design, expression, and purification of an active C-terminal polymerase fragment. Strategies to obtain and improve crystals of a ternary POLQ complex (enzyme:DNA:nucleotide) are also presented, along with key elements of the structure. PMID: 28668117
Our results indicate that there is a synthetic lethal relationship between pol theta;-mediated DNA repair and homologous recombination pathways PMID: 27533083
Data suggest that error-free DNA replication through 3-deaza-3-methyladenine adduct is mediated via three different pathways dependent upon POL-iota/POL-kappa, POL-theta, and POL-zeta. PMID: 28939775
Use fluorescence resonance energy transfer to monitor assembly of the human replicative polymerase holoenzyme. PMID: 23577232
These results suggest that variants in the POLQ gene may be associated with the risk of Luminal breast cancer PMID: 25417172
A DNA repair variant in POLQ (c.-1060A > G) is associated to hereditary breast cancer patients. PMID: 25409685
Polymerase theta; uses a specialized thumb subdomain to establish unique upstream contacts to the primer DNA strand. PMID: 25775267
Microhomology-mediated end-joining is dependent on Poltheta; in human cells. PMID: 25643323
depletion of Poltheta; has a synergistic effect on cell survival in the absence of BRCA genes, suggesting that the inhibition of this mutagenic polymerase represents a valid therapeutic avenue for tumours carrying mutations in homology-directed repair genes PMID: 25642960
results reveal a synthetic lethal relationship between the homologous-recombination pathway and Poltheta;-mediated repair in epithelial ovarian cancers, and identify Poltheta; as a novel druggable target for cancer therapy PMID: 25642963
A role for DNA polymerase theta; in promoting replication through oxidative DNA lesion, thymine glycol, in human cells. PMID: 24648516
POLQ possesses a DNA polymerase activity that appears to be template independent and allows efficient extension of single-stranded DNA as well as duplex DNA with either protruding or multiply mismatched 3'-OH termini. PMID: 22135286
overexpression in breast cancer confers an adverse prognosis and is associated with key cancer pathways PMID: 20700469
Data show that POLQ overexpression may be a promising genetic instability and prognostic marker for breast cancer. PMID: 20624954
DNA polymerase theta purified from human cells is a high-fidelity enzyme. PMID: 12051913
the isolation of the full-length human DNA POLQ gene, and an initial characterization of its gene product, DNA polymerase theta PMID: 14576298
DNA Pol theta has a specialized function in lymphocytes and in tumor progression PMID: 14735462
POLQ has a high efficiency in by-passing DNA damage. PMID: 15496986
analysis of human DNA polymerase eta error-prone synthesis on DNA deoxyadenosine adducts PMID: 16188888
The results demonstrate that activation of a UV-induced DNA damage response pathway, involving phosphorylation of RPA p34 by DNA-PK, is enhanced in cells lacking poleta. PMID: 16520097
Pol eta can play an important role in determining the cellular sensitivity to therapeutic agents. PMID: 16603639
DNA polymerase eta (Poleta) is responsible for efficient translesion synthesis (TLS) past cis-syn cyclobutane thymine dimers (TT dimers), the major DNA lesions induced by UV irradiation. PMID: 16824193
Human DNA polymerase eta selectively produces a two-base deletion in copying the N2-guanyl adduct of 2-amino-3-methylimidazo[4,5-f]quinoline but not the C8 adduct at the NarI G3 site PMID: 16835218
The enzymatic reactions with human DNA polymerase eta on oxidative products of guanine and 8-oxoG, is investigated. PMID: 17150533
Evolutionary conservation of efficient T[CPD]T bypass by HsPoleta and AtPoleta may reflect a high degree of exposure of human skin and plants to solar UV-B radiation PMID: 18366182
When copying undamaged DNA, DNA polymerase theta generates single base errors at rates 10- to more than 100-fold higher than for other family A members. PMID: 18503084
Domain mapping of the 98-kDa enzyme by limited proteolysis and NaBH(4) cross-linking with a base excision repair intermediate revealed that the dRP lyase active site resides in a 24-kDa domain of Pol theta. PMID: 19188258

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Involvement in disease

Breast cancer (BC)

Subcellular Location

Nucleus. Chromosome.

Protein Families

DNA polymerase type-A family

Tissue Specificity

Highly expressed in testis.

Database Links

HGNC: 9186

OMIM: 114480

KEGG: hsa:10721

STRING: 9606.ENSP00000264233

UniGene: Hs.241517

Code	CSB-EP018321HU
Abbreviation	Recombinant Human POLQ protein, partial
MSDS	MSDS Datasheet
Size	$306
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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 Human DNA polymerase theta (POLQ), partial

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