Recombinant Ceratophyllum demersum Apocytochrome f (petA)

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Cat. No.
BT2478253
Source
in vitro E.coli expression system
Synonyms
petA; Cytochrome f
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Product Specs

Form
Lyophilized powder.
Note: While we prioritize shipping the format currently in stock, please specify your format preference during order placement for customized preparation.
Lead Time
Delivery times vary depending on the purchasing method and location. Please consult your local distributor for precise delivery estimates.
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Notes
Avoid repeated freeze-thaw cycles. Store working aliquots at 4°C for up to one week.
Reconstitution
Centrifuge the vial briefly before opening to collect the contents. Reconstitute the protein in sterile, deionized water to a concentration of 0.1-1.0 mg/mL. We recommend adding 5-50% glycerol (final concentration) and aliquoting for long-term storage at -20°C/-80°C. Our default glycerol concentration is 50% and can serve as a guideline.
Shelf Life
Shelf life depends on storage conditions, buffer components, temperature, and protein stability. Generally, liquid formulations have a 6-month shelf life at -20°C/-80°C, while lyophilized forms have a 12-month shelf life at -20°C/-80°C.
Storage Condition
Upon receipt, store at -20°C/-80°C. Aliquoting is essential for multiple uses. Avoid repeated freeze-thaw cycles.
Tag Info
Tag type is determined during manufacturing.
The tag type will be determined during the production process. If you require a specific tag type, please inform us, and we will prioritize its development.
Synonyms
petA; Cytochrome f
Buffer Before Lyophilization
Tris/PBS-based buffer, 6% Trehalose.
Datasheet
Please contact us to get it.
Expression Region
36-320
Protein Length
Full Length of Mature Protein
Species
Ceratophyllum demersum (Rigid hornwort) (Coontail)
Target Names
petA
Target Protein Sequence
YPIFAQQGYENPREATGRIVCANCHLANKPVDIEVPQAVLPDTVFEAVVRIPYDMQLKQV LANGKKGGLNVGAVLILPEGFELAPPDRISPEMKEKIGNLSFQSYRPNKKNILIMGPVPG KKYSQIVFPILSPDPATKKDVHFLKYPIYVGGNRGRGQIYPDGSKSNNTVYNATATGIVS KIVRKEKGGYEITIADTADGHQVVDIIPPGPELLVSEGESIKIDQPLTSNPNVGGFGQGD AEIVLQDPLRVQGLLFFFASVILAQIFLVLKKKQFEKVQLSEMNF
Uniprot No.
A8SEB6

Target Background

Function
A component of the cytochrome b6-f complex, mediating electron transfer between Photosystem II (PSII) and Photosystem I (PSI), cyclic electron flow around PSI, and state transitions.
Protein Families
Cytochrome f family
Subcellular Location
Plastid, chloroplast thylakoid membrane; Single-pass membrane protein.

Q&A

FAQs for Researchers on Recombinant Ceratophyllum demersum Apocytochrome f (petA)

Advanced Research Questions

  • How can researchers resolve discrepancies in recombinant petA activity assays?
    Common issues and solutions:

    • Low electron transfer rates: Verify heme incorporation via UV-Vis spectroscopy (peak at 553 nm) .

    • Aggregation during purification: Use zwitterionic detergents (e.g., LDAO) in lysis buffers .

    • Inconsistent complementation in Arabidopsis mutants: Ensure proper chloroplast targeting by adding transit peptides (e.g., from RbcS) in transgenic constructs .

  • What strategies enhance recombinant petA stability for structural studies?

    • Lyophilization: Store in Tris/PBS buffer with 6% trehalose (pH 8.0) to prevent denaturation .

    • Crystallization additives: Include 0.1% n-dodecyl-β-D-maltoside (DDM) and 10 mM ascorbate to maintain redox state .

  • How does petA interact with heavy metal detoxification pathways in transgenic plants?
    Co-expression with C. demersum phytochelatin synthase (CdPCS1) in tobacco increases cadmium (Cd) and arsenic (As) accumulation by 3–5×. This synergy arises from petA’s role in reducing oxidative stress, enhancing phytochelatin-mediated metal sequestration .

    Transgenic LineCd Accumulation (μg/g DW)As Accumulation (μg/g DW)
    Wild-type tobacco12.5 ± 1.28.3 ± 0.9
    petA + CdPCS158.7 ± 3.541.2 ± 2.8

  • What bioinformatics tools are critical for analyzing petA evolutionary conservation?

    • Phylogenetic placement: Use RAxML with plastome datasets (e.g., 75 protein-coding genes) .

    • Homology modeling: SWISS-MODEL with templates like Spinacia oleracea (PDB: 1Q90) .

    • Codon optimization: JCat for adapting petA to host organisms (e.g., tobacco codon usage) .

Methodological Guidance

  • Troubleshooting low yields: Screen induction temperatures (16°C, 25°C) and use E. coli C43(DE3) for membrane proteins .

  • Validating function: Perform in vitro reconstitution with ferredoxin-NADP+ reductase (FNR) to assay electron transfer rates .

  • Ethical considerations: Avoid human consumption due to potential allergenicity from bacterial endotoxins .

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