Recombinant Escherichia coli CRISPR-associated helicase Cas3 (ygcB), partial

Code CSB-YP334357ENV
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Source Yeast
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Code CSB-EP334357ENV
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Source E.coli
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Code CSB-EP334357ENV-B
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Source E.coli
Conjugate Avi-tag Biotinylated
E. coli biotin ligase (BirA) is highly specific in covalently attaching biotin to the 15 amino acid AviTag peptide. This recombinant protein was biotinylated in vivo by AviTag-BirA technology, which method is BriA catalyzes amide linkage between the biotin and the specific lysine of the AviTag.
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Code CSB-BP334357ENV
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Source Baculovirus
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Code CSB-MP334357ENV
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Source Mammalian cell
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Product Details

Purity
>85% (SDS-PAGE)
Target Names
ygcB
Uniprot No.
Alternative Names
ygcB; cas3; b2761; JW2731; CRISPR-associated endonuclease/helicase Cas3; EC 3.1.-.-; EC 3.6.4.-
Species
Escherichia coli (strain K12)
Protein Length
Partial
Tag Info
Tag type will be determined during the manufacturing process.
The tag type will be determined during production process. If you have specified tag type, please tell us and we will develop the specified tag preferentially.
Form
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 before Lyophilization
Tris/PBS-based buffer, 6% Trehalose, pH 8.0
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℃/-80℃. Our default final concentration of glycerol is 50%. Customers could use it as reference.
Troubleshooting and 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
Delivery time may differ from different purchasing way or location, please kindly consult your local distributors for specific delivery time.
Note: All of our proteins are default shipped with normal blue ice packs, if you request to ship with dry ice, please communicate with us in advance and extra fees will be charged.
Notes
Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.
Datasheet
Please contact us to get it.

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Target Background

Function
CRISPR (clustered regularly interspaced short palindromic repeat), is an adaptive immune system that provides protection against mobile genetic elements (viruses, transposable elements and conjugative plasmids). CRISPR clusters contain sequences complementary to antecedent mobile elements and target invading nucleic acids. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA). Cas3 plus Cascade participate in CRISPR interference, the third stage of CRISPR immunity.; Acts as an endonuclease, a 3'-5'exonuclease, and an ATP-dependent dsDNA helicase. Anneals and unwinds R-loops (in which crRNA binds the target DNA, displacing the noncomplementary strand). Unwinding requires ATP, annealing does not. Required along with the Cascade complex for resistance to bacteriophage lambda infection as well as the ability to cure CRISPR-encoding high-copy number plasmid. A Cas3-CasA fusion protein purified with the Cascade complex nicks target plasmid in the presence but not absence of Mg(2+), and degrades plasmid fully in the presence of Mg(2+) and ATP, suggesting the helicase activity is required for complete degradation.
Gene References into Functions
  1. Cas3-derived target DNA degradation fragments fuel primed CRISPR adaptation. PMID: 27546790
  2. study observed that Cas3 promoted runaway ColE1 plasmid replication, requiring Cas3 helicase activity; Cas3 expression antagonized RNaseHI, observed as a concatamer phenotype, but also required RNaseHI for its ability to stimulate plasmid copy number PMID: 23406879
  3. Cas3 is involved in DNA degradation and invader neutralization. PMID: 22521689
  4. Study discuss possible overlapping roles of Cas3 and Cascade in E. coli, and the apparently antagonistic roles of Cas3 catalysing RNA-DNA annealing and ATP-dependent helicase unwinding. PMID: 21699496
Protein Families
CRISPR-associated nuclease Cas3-HD family; CRISPR-associated helicase Cas3 family
Database Links
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