🔬 Research Methodology

Step 1: Gene Optimization

Identify optimal pcrA/pcrB and cld genes, then codon-optimize for B. subtilis using JCat and GeneOptimizer.

Step 2: Pathway Modeling

Integrate perchlorate detox pathway into B. subtilis metabolic network and simulate Mars-like conditions.

Step 3: Enzyme Engineering

Dock enzymes with perchlorate/chlorite and run molecular dynamics to confirm cold-environment stability.

Step 4: Genome Engineering

Design plasmids or apply CRISPR editing for chromosomal integration using strong promoters.

Step 5: Validation

RNA-Seq and WGS to confirm expression, stability, and check for off-target effects.

Step 6: Mars Simulation

Model system performance in low-pressure, low-temperature environments with ML optimization.

Step 7: Bioethics & Safety

Align with NASA Planetary Protection guidelines and address biosafety concerns.