Recombinant Guillardia theta ATP-dependent zinc metalloprotease FtsH (ftsH)

What is the molecular architecture of Guillardia theta FtsH and how does it function?

The ATP-dependent integral membrane protease FtsH from Guillardia theta forms a complex hexameric structure essential for its function. The molecular architecture consists of two rings where the protease domains possess an all-helical fold and form a flat hexagon covered by a toroid built by the AAA domains. The active site classifies FtsH as an Asp-zincin metalloprotease.

Key structural elements include:

• Two histidine residues (His423 and His427) in the conserved HEXXH motif serve as zinc ligands

• The third zinc ligand is Asp-500 (not Glu-486 as previously reported)

• The different symmetries of protease and AAA rings suggest a translocation mechanism for target polypeptides

FtsH functions through ATP-dependent proteolysis, where the AAA domain provides energy for substrate unfolding and translocation into the proteolytic chamber. This is essential for the degradation of damaged or unneeded membrane proteins, playing a crucial role in protein quality control .

How do the transmembrane domains contribute to FtsH function?

The transmembrane helices, particularly the second one, are essential for three key aspects of FtsH function:

• Oligomerization: They facilitate the formation of the hexameric FtsH complex

• ATPase activity: They contribute to proper positioning for efficient ATP hydrolysis

• Proteolytic activity: They help maintain the structural integrity needed for optimal proteolysis

Electron microscopic studies have confirmed that FtsH forms ring-shaped toroidal oligomers like other AAA proteins. While a hexameric structure has been predicted based on comparison with other AAA domains, the transmembrane domains play a critical role in stabilizing this quaternary structure in addition to anchoring the protein in the membrane .

What are the recommended assays for measuring recombinant FtsH proteolytic activity?

For reliable measurement of recombinant Guillardia theta FtsH proteolytic activity, the following methodology has been established:

Beta-casein degradation assay:

• Beta-casein serves as an ideal substrate due to its lack of defined secondary and tertiary structures

• The reaction mixture should contain purified recombinant FtsH, beta-casein, and ATP

• Degradation can be detected using Coomassie blue staining of SDS-PAGE gels

Controls and validation:

• Include compact globular proteins (BSA, GST) as negative controls that should remain stable throughout the experiment

• Include reactions without ATP to confirm ATP dependency

• Test inhibitor specificity: o-phenanthroline (a metalloprotease inhibitor) should inhibit activity, while serine and cysteine protease inhibitors should not affect caseinolytic activity

D1 fragment degradation:

• For more physiologically relevant assays, the 23-kD fragment of photosystem II D1 protein can be used as a substrate

• This degradation is dependent on ATP hydrolysis and divalent metal ions, confirming FtsH involvement .

What expression and purification strategies yield functional recombinant FtsH protein?

Expression Systems:

• E. coli is the most commonly used expression host for recombinant Guillardia theta FtsH

• The full-length protein (residues 1-631) can be successfully expressed with N-terminal His-tags

Purification Method:

• Immobilized metal affinity chromatography (IMAC) using the His-tag

• Size exclusion chromatography to isolate properly folded hexameric complexes

Buffer Considerations:

• Include zinc ions to maintain metalloprotease activity

• ATP and Mg²⁺ are essential for proper folding and activity

• Avoid chelating agents that may sequester the zinc ions

Storage Conditions:

• Store at -20°C/-80°C for extended storage

• Maintain working aliquots at 4°C for up to one week

• Avoid repeated freeze-thaw cycles which can diminish activity

Lyophilized preparations with 6% trehalose at pH 8.0 have been shown to maintain protein stability .

How can mutations in the active site be designed to study FtsH mechanism?

To investigate the catalytic mechanism of FtsH metalloprotease activity, strategic mutations can be designed targeting key residues in the active site:

Critical Residues for Mutagenesis:

• HEXXH Motif (residues 423-427): Mutating either histidine disrupts zinc coordination

• Asp-500: The D500A mutation completely abolishes proteolytic activity, confirming its role as the third zinc ligand

• Glu-486: E486V mutation retains approximately 10% activity, as it forms hydrogen bonds to stabilize the imidazole side chain of the first histidine

Experimental Design Strategy:

• Generate single point mutations using site-directed mutagenesis

• Express and purify mutant proteins using the same protocol as wild-type

• Compare proteolytic activity against standard substrates (e.g., beta-casein)

• Perform structural analysis to confirm altered zinc coordination

Crystal structure analysis of the D500A mutant confirmed the loss of the zinc ion, validating its essential role in FtsH catalysis .

What is the role of FtsH in photosystem II repair?

FtsH plays a critical role in the quality control of photosystem II, particularly in the turnover of the D1 reaction center protein:

D1 Protein Turnover Mechanism:

• The D1 protein is damaged by reactive oxygen species formed during photosynthesis

• The damaged D1 protein undergoes primary cleavage, generating a 23-kD fragment

• FtsH degrades this fragment in an ATP and divalent metal ion-dependent process

• A newly synthesized D1 protein replaces the degraded one

Experimental Evidence:

• Purified FtsH degrades the 23-kD D1 fragment in isolated photosystem II core complexes

• This degradation occurs in thylakoid membranes depleted of endogenous FtsH

• The process is dependent on ATP hydrolysis and divalent metal ions

• The 23-kD fragment is located in stroma-exposed thylakoid regions, accessible to FtsH

This repair cycle is essential for maintaining photosynthetic efficiency under varying light conditions and preventing photoinhibition .

How does FtsH function relate to leaf variegation in plants?

The relationship between FtsH and leaf variegation has been extensively studied in Arabidopsis thaliana mutants:

Key Findings:

• The leaf-variegated mutant yellow variegated2 (var2) results from loss of FtsH2

• Similarly, var1 results from the loss of FtsH5

• Both FtsH2 and FtsH5 are major components of the chloroplast FtsH complex

• The variegated phenotype is specific to these two FtsH proteins; knockouts of other FtsH genes do not show visible phenotypes

FtsH Complex Composition:

• FtsH2 and FtsH5 form a major heterocomplex of ~400 kD

• There are two types of chloroplast FtsHs:

  • Type A: FtsH1/FtsH5

  • Type B: FtsH2/FtsH8

• Both types are functionally distinguishable and required for proper chloroplast development

Suppression Mechanism:

• Second-site mutations like fu-gaeri1 (fug1) can suppress leaf variegation

• fug1 encodes a protein homologous to prokaryotic translation initiation factor 2 (cpIF2)

• Reduced chloroplast translation can suppress the variegated phenotype

This suggests that the balance between protein synthesis and degradation is a critical factor in chloroplast maintenance and thylakoid development .

What is known about FtsH gene diversity and expression across plant species?

Recent genomic studies have revealed significant diversity in the FtsH gene family across plant species:

Wheat FtsH Family Characteristics:

• 11 TaFtsH genes identified with uneven chromosomal distribution

• Significant variations in gene sequence length and intron numbers

• Classified into eight groups with similar structures and conserved motifs

• Extensive gene duplications within the TaFtsH gene family

• Closer relationship to maize FtsH genes than other species

Expression Patterns:

• TaFtsH genes are expressed in all wheat tissues but with varying patterns

• Differential responses to metal stress treatments (CdCl₂, ZnSO₄, and MnSO₄)

• Under CdCl₂ stress, expression levels of TaFtsH-1, TaFtsH-2, TaFtsH-5, TaFtsH-9, and TaFtsH-11 peaked at 24 hours

• Under ZnSO₄ stress, 5 of 11 TaFtsH genes were generally upregulated over time

• Under MnSO₄ treatment, several genes increased and then decreased after 12 hours

Functional Significance:

• Gene Ontology (GO) enrichment analysis indicates TaFtsH genes are involved in protein hydrolysis

• Silencing TaFtsH-1 enhances wheat's resistance to cadmium toxicity

• Developmental and stress-responsive elements were found in the promoter regions of most TaFtsH genes

This diversity suggests specialized roles for different FtsH proteins in plant development and stress responses .

How do the symmetry mismatches between AAA and protease domains impact FtsH function?

The molecular architecture of FtsH reveals an intriguing asymmetry between its functional domains:

Observed Symmetry Mismatch:

• The protease domains form a flat hexagon with six-fold symmetry

• The AAA ring shows a breakdown of the expected hexagonal symmetry to C2 symmetry

Functional Implications:

• This symmetry mismatch may be essential for the catalytic cycle

• Similar to the symmetry mismatch between hexameric ClpX ATPase and heptameric ClpP protease in the ClpXP complex

• The distortion from C6 to C2 symmetry resembles that observed in T7 gene 4 ring helicase

Mechanistic Model:

• The symmetry reduction likely facilitates sequential nucleotide hydrolysis

• This creates a coordinated "power stroke" for substrate translocation into the proteolytic chamber

• The asymmetric conformation allows for coordinated binding, hydrolysis, and release of ATP

• This mechanism optimizes the energy usage for polypeptide unfolding and translocation

This structural arrangement suggests a sophisticated molecular machine that converts ATP hydrolysis into mechanical force for protein degradation .

What strategies can be used to study FtsH interactions with membrane proteins in their native environment?

Investigating FtsH interactions with membrane proteins presents unique challenges that require specialized approaches:

Recommended Methodologies:

• Crosslinking Mass Spectrometry (XL-MS)

  • Chemical crosslinkers can capture transient interactions between FtsH and substrates

  • Mass spectrometry identifies the crosslinked peptides

  • Provides spatial constraints for modeling protein-protein interactions

• Reconstituted Proteoliposome Systems

  • Incorporate purified recombinant FtsH into liposomes

  • Add potential substrate proteins to the liposomes

  • Monitor degradation using proteomics approaches

• Thylakoid Membrane Isolation

  • Isolate intact thylakoid membranes containing endogenous FtsH

  • Deplete endogenous FtsH and reconstitute with recombinant variants

  • Study degradation of photosystem components like the D1 protein

• Fluorescence Resonance Energy Transfer (FRET)

  • Label FtsH and potential substrates with fluorescent tags

  • Monitor interactions through changes in FRET efficiency

  • Can be performed in isolated membranes or reconstituted systems

• In vivo Proximity Labeling

  • Fuse FtsH to enzymes like BioID or APEX2

  • These enzymes biotinylate proximal proteins upon activation

  • Identify labeled proteins by streptavidin pull-down and mass spectrometry

These approaches can reveal the molecular mechanisms of substrate recognition and processing by FtsH in its native membrane environment .

How might the study of Guillardia theta FtsH inform our understanding of secondary endosymbiosis?

Guillardia theta, as a cryptomonad containing a nucleomorph genome, offers unique insights into secondary endosymbiosis processes:

Evolutionary Context:

• G. theta contains a plastid derived from a red algal endosymbiont

• The nucleomorph is the highly reduced nucleus of the engulfed red alga

• The chloroplast genome of G. theta has unique characteristics compared to red algae

Structural Evidence:

• The G. theta chloroplast genome contains inverted repeats, unlike the direct repeats found in red algae

• These inverted repeats evolved after secondary endosymbiosis through recombination of rRNA cistrons

• This suggests distinct evolutionary processes during endosymbiont integration

Comparative Analysis:

• Nucleomorph genome size in Hemiselmis (another cryptomonad genus) ranges from 560 to 600 kb

• Different Hemiselmis species show distinct nucleomorph genome karyotypes

• Nucleomorph-encoded genes, including those potentially interacting with FtsH, provide insights into the redistribution of genetic material during endosymbiont reduction

Gene Transfer Implications:

• FtsH in G. theta may represent a case of functional gene transfer between endosymbiont and host

• Studying FtsH localization and function in cryptomonads can reveal mechanisms of protein targeting and integration during endosymbiosis

• The coordination between nuclear-encoded and nucleomorph-encoded proteins in maintaining the chromalveolate plastid

These insights contribute to our understanding of the evolutionary processes that shape endosymbiotic relationships and organelle evolution .

What are common challenges in maintaining FtsH activity during recombinant protein preparation?

Several technical challenges can affect the successful preparation of active recombinant Guillardia theta FtsH:

Challenge 1: Maintaining Zinc Coordination

• As a zinc metalloprotease, FtsH requires proper zinc coordination for activity

• Solutions:

  • Include zinc ions (1-10 μM ZnCl₂) in purification buffers

  • Avoid chelating agents like EDTA

  • Use o-phenanthroline as a control inhibitor to confirm zinc-dependent activity

Challenge 2: ATP-Dependent Activity

• FtsH requires ATP for both structural integrity and proteolytic function

• Solutions:

  • Include ATP (1-5 mM) and Mg²⁺ (5-10 mM) in activity assays

  • For long-term storage, consider ATP analogs that resist hydrolysis

  • Verify ATP dependency by comparing activity with and without ATP

Challenge 3: Protein Stability

• Recombinant FtsH can lose activity during storage and freeze-thaw cycles

• Solutions:

  • Store at -20°C/-80°C with 50% glycerol or 6% trehalose

  • Prepare single-use aliquots to avoid repeated freeze-thaw cycles

  • Keep working aliquots at 4°C for no more than one week

Challenge 4: Oligomerization

• Functional FtsH requires proper hexameric assembly

• Solutions:

  • Verify oligomeric state using size exclusion chromatography

  • Consider mild detergents to maintain membrane protein solubility

  • Include stabilizing agents like glycerol or non-detergent sulfobetaines

Challenge 5: Expression Construct Design

• Full-length versus truncated constructs affects activity and solubility

• Solutions:

  • For functional studies, use constructs with intact AAA and protease domains

  • Test multiple constructs with different boundaries around transmembrane regions

  • Consider fusion proteins or solubility tags that can be later removed .

How can researchers distinguish between the activities of different FtsH homologs in complex biological samples?

Differentiating between various FtsH homologs in biological samples requires strategic experimental approaches:

Methodological Approaches:

• Gene-Specific Knockdown/Knockout

  • Use CRISPR-Cas9 or RNAi to target specific FtsH genes

  • BSMV-VIGS technology has been successfully used for silencing TaFtsH-1 in wheat

  • Compare phenotypes and molecular profiles before and after targeted manipulation

• Antibody-Based Detection

  • Develop antibodies against unique epitopes of specific FtsH proteins

  • Use these for immunoblot analysis and immunoprecipitation

  • Example: antibodies against cpIF2 successfully distinguished between FtsH variants in Arabidopsis

• Substrate Specificity Analysis

  • Different FtsH homologs may have distinct substrate preferences

  • Design substrate competition assays to identify preferential degradation

  • Monitor degradation kinetics using fluorescent or tagged substrates

• Expression Pattern Analysis

  • RT-qPCR with gene-specific primers can distinguish expression patterns

  • Different FtsH genes show distinct responses to stressors

  • For example, under CdCl₂ stress, expression levels of TaFtsH-1, TaFtsH-2, TaFtsH-5, TaFtsH-9, and TaFtsH-11 peaked at 24 hours

• Comparative Proteomics

  • Mass spectrometry can identify specific FtsH proteins in complex samples

  • Targeted proteomics (PRM/MRM) can quantify specific peptides unique to each FtsH variant

  • Stable isotope labeling can track turnover rates of different FtsH proteins

These approaches allow researchers to distinguish between the activities and functions of different FtsH homologs in complex biological systems .

What regulatory compliance considerations apply when working with recombinant FtsH in research settings?

When working with recombinant Guillardia theta FtsH in research laboratories, several regulatory compliance considerations must be addressed:

NIH Guidelines Compliance:

• Institutions receiving NIH funding must follow the NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules

• Even if only one project at an institution receives NIH funding, all recombinant DNA projects must comply

• Other funding agencies may also require adherence to NIH Guidelines

Institutional Biosafety Committee (IBC) Review:

• Research involving recombinant DNA requires IBC review and approval

• The IBC assesses biosafety risks and determines appropriate containment measures

• Experiments involving more than 10 liters of culture may require additional oversight

Risk Assessment Factors:

• Expression system (E. coli, yeast, baculovirus, or mammalian cells)

• Vector characteristics (mobilization potential, antibiotic resistance markers)

• Insert characteristics (FtsH itself is not a toxin or virulence factor)

• Scale of operation (large-scale work over 10 liters requires additional considerations)

Documentation Requirements:

• Detailed experimental protocols

• Risk assessment documentation

• Training records for laboratory personnel

• Records of IBC approval and any stipulations

For most recombinant FtsH work in standard research settings, Biosafety Level 1 (BSL-1) practices are typically sufficient when using non-pathogenic expression systems, but institutional guidelines should always be consulted for specific requirements .

How might emerging single-molecule techniques advance our understanding of FtsH mechanism?

Single-molecule techniques offer exciting opportunities to investigate FtsH dynamics that are obscured in bulk measurements:

Promising Techniques:

• Single-Molecule FRET (smFRET)

  • Can track conformational changes during ATP hydrolysis and substrate processing

  • Allows visualization of the hexameric ring dynamics during substrate translocation

  • Could reveal asymmetric conformational changes implied by the symmetry mismatch

• Optical Tweezers

  • Can measure the force generated by FtsH during substrate unfolding and translocation

  • May identify the step size of substrate movement through the proteolytic chamber

  • Could quantify how ATP hydrolysis is coupled to mechanical work

• Single-Molecule Fluorescence Microscopy

  • Track the movement of fluorescently labeled substrates through FtsH

  • Monitor real-time degradation of individual substrate molecules

  • Identify potential stochastic behaviors in the degradation process

• High-Speed Atomic Force Microscopy (HS-AFM)

  • Directly visualize conformational changes in the FtsH hexamer during function

  • Observe substrate binding and translocation in real-time

  • Map structural dynamics across the AAA and protease domains

• Nanopore Technology

  • Engineer FtsH to function as a biological nanopore

  • Monitor substrate translocation through current fluctuations

  • Provide insights into the selectivity and mechanism of substrate processing

These approaches could transform our understanding of how FtsH converts ATP hydrolysis into mechanical force for substrate translocation and degradation, potentially revealing unknown intermediate states during the reaction cycle .

What can comparative genomics of FtsH across different algal lineages tell us about evolutionary adaptation?

Comparative genomic analysis of FtsH across algal lineages offers rich insights into evolutionary adaptation:

Research Opportunities:

• Diversification Patterns

  • Compare FtsH gene families across primary and secondary endosymbiotic algae

  • Investigate whether similar duplications and specializations occurred independently

  • Analyze selection pressures across different algal habitats

• Secondary Endosymbiosis Events

  • Compare FtsH from different lineages resulting from independent secondary endosymbiosis

  • Investigate convergent or divergent evolutionary paths in different cryptophytes, haptophytes, and stramenopiles

  • Determine whether similar targeting mechanisms evolved independently

• Extremophile Adaptations

  • Study FtsH in thermophilic algae like Cyanidioschyzon caldarium

  • Investigate structural adaptations for stability under extreme conditions

  • Explore potential specialized roles in stress resistance

• Gene Transfer Patterns

  • Track the migration of FtsH genes between endosymbiont and host genomes

  • Identify key transitions in targeting peptide acquisition

  • Map the evolutionary history of regulatory elements controlling FtsH expression

• Coevolution with Photosystems

  • Analyze how FtsH evolution correlates with changes in photosystem components

  • Investigate whether FtsH adaptations track with specific photosynthetic strategies

  • Study coevolution patterns between FtsH and its substrate proteins

Full-length transcriptome analysis technologies, such as those applied to Akashiwo sanguinea and other algae, have enhanced our ability to identify and characterize FtsH genes across diverse lineages, opening new avenues for evolutionary research .

How might engineered variants of FtsH be utilized in biotechnology applications?

Engineered variants of FtsH hold significant potential for diverse biotechnology applications:

Potential Applications:

• Protein Quality Control in Recombinant Production

  • Engineer FtsH to selectively degrade misfolded proteins in expression systems

  • Improve yield and quality of difficult-to-express proteins

  • Design substrate-specific variants for targeted degradation

• Stress-Resistant Crop Development

  • Introduce modified FtsH variants into crop plants to enhance stress tolerance

  • Building on findings that silencing TaFtsH-1 enhances cadmium resistance in wheat

  • Develop plants with improved photosystem repair under high light or heavy metal stress

• Synthetic Biology Tools

  • Create inducible protein degradation systems based on FtsH

  • Develop orthogonal protein quality control systems for synthetic cells

  • Engineer conditional protein knockout systems based on FtsH recognition

• Bioremediation

  • Design FtsH variants that enhance algal survival in heavy metal-contaminated waters

  • Develop engineered algae with improved metal sequestration capabilities

  • Create biological systems for environmental cleanup with regulated protein turnover

• Therapeutic Applications

  • Target human FtsH homologs (e.g., those implicated in spastic paraplegia)

  • Develop small molecule modulators of FtsH activity for disease treatment

  • Engineer therapeutic cells with enhanced stress resistance via modified FtsH activity

These applications leverage the fundamental understanding of FtsH structure-function relationships to create novel biotechnological tools and strategies, highlighting the translation of basic research into practical applications .

What are the key considerations when selecting commercial recombinant FtsH for research applications?

When selecting commercial recombinant Guillardia theta FtsH for research applications, consider these critical factors:

Product Specifications to Evaluate:

• Protein Length and Region

  • Full-length protein (1-631 amino acids) versus partial constructs

  • Presence or absence of transmembrane domains affects application suitability

  • Full-length proteins are better for structural studies but may have solubility challenges

• Expression System

  • E. coli-expressed proteins are most common and economical

  • Eukaryotic expression systems (yeast, insect cells) may provide better folding

  • Consider potential endotoxin contamination from bacterial systems if using for cell culture

• Purification Tags

  • N-terminal or C-terminal His-tags are most common (10xHis or 6xHis)

  • Tag location may affect activity or accessibility of certain domains

  • Consider whether tag removal is necessary for your application

• Purity and Quality Control

  • Verify SDS-PAGE purity (typically ≥85-90%)

  • Request lot-specific quality control data

  • Check for functional validation (ATPase or proteolytic activity)

• Formulation and Storage

  • Lyophilized versus liquid formulations affect stability and convenience

  • Buffer composition (presence of glycerol, trehalose, or other stabilizers)

  • Storage requirements (-20°C/-80°C) and shelf life

• Supporting Documentation

  • Certificate of Analysis with lot-specific data

  • Technical datasheet with handling instructions

  • MSDS for safety information

Several suppliers offer recombinant Guillardia theta FtsH with prices typically around $2,000-2,200 per unit (50 μg) .

What sequence databases and bioinformatic tools are most useful for FtsH research?

Researchers studying FtsH can benefit from these specialized databases and bioinformatic tools:

Sequence Databases:

• UniProt - Contains curated FtsH sequences (e.g., Guillardia theta FtsH: O78516)

• NCBI Protein Database - Comprehensive collection of protein sequences including numerous FtsH homologs

• Plant TFdb - Useful for analyzing transcription factors that regulate FtsH expression

• ChloroP and TargetP - For prediction of chloroplast transit peptides in FtsH proteins

Structural Resources:

• Protein Data Bank (PDB) - Contains crystal structures of FtsH domains

• ModBase - Provides 3D structure models for proteins like FtsH (referenced for O78516)

• AAA+ Database - Specialized resource for AAA+ protein family members including FtsH

Analysis Tools:

• MISA (Microsatellite Identification Tool) - For identifying simple sequence repeats within FtsH genes

• KEGG Database - For mapping FtsH in metabolic and signaling pathways

• Gene Ontology (GO) - For functional annotation of FtsH proteins

• SUPPA - For detection of alternative splicing events in FtsH transcripts

• Cogent (Coding GENome reconstruction Tool) - For analyzing transcript isoforms

Expression Data Resources:

• SIGnAL T-DNA Express Database - For identifying T-DNA insertion mutants in model organisms

• RNA-Seq Expression Browsers - For tissue-specific expression patterns of FtsH genes

These resources facilitate comparative analysis, functional prediction, and evolutionary studies of FtsH proteins across diverse organisms .

What model systems are most suitable for studying FtsH function in vivo?

Several model systems offer distinct advantages for studying FtsH function in vivo:

Plant Models:

• Arabidopsis thaliana

  • Well-characterized var1 and var2 mutants lacking FtsH5 and FtsH2, respectively

  • Extensive genetic tools and mutant collections

  • Clearly visible leaf variegation phenotype for monitoring FtsH function

  • Second-site suppressors like fug1 provide insights into functional networks

• Wheat (Triticum aestivum)

  • 11 identified TaFtsH genes with diverse expression patterns

  • BSMV-VIGS technology effective for gene silencing

  • Agriculturally relevant for studying stress responses

  • Polyploid genome allows study of gene redundancy and specialization

Algal Models:

• Guillardia theta

  • Natural source of the FtsH studied

  • Provides insights into secondary endosymbiosis

  • Nucleomorph genome offers unique evolutionary perspective

  • Full protein sequence and recombinant protein commercially available

• Chlamydomonas reinhardtii

  • Well-established genetic tools for chloroplast biology

  • Haploid genome simplifies genetic manipulation

  • Rapid growth and well-characterized photosynthetic apparatus

Cyanobacterial Model:

• Synechocystis sp. PCC 6803

  • Prokaryotic system with simplified genetic background

  • Photosynthetic capabilities similar to chloroplasts

  • Established transformation protocols

  • Rapid growth and relatively simple genome

Each system offers unique advantages depending on the specific research question, from basic mechanistic studies to applied agricultural applications .