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Revision as of 11:52, 16 October 2018

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Parts

Basic Parts

BBa_K2567000: nicA (Nicotine oxidoreductase)

The part code for the nicA protein. This protein can degrade nicotine. Nicotine oxidoreductase (nicA) from Pseudomonas putida S16 is the first enzyme in the pyrrolidine pathway for nicotine degradation.

We optimization the nicA in order to express protein successfully. Including but not limited to:

• Codon usage bias

• GC content

• CpG dinucleotides content

• mRNA secondary structure

• Cryptic splicing sites

• Premature PolyA sites

• Internal chi sites and ribosomal binding sites

• Negative CpG islands

• RNA instability motif (ARE)

• Repeat sequences (direct repeat, reverse repeat, and Dyad repeat)

• Restriction sites that may interfere with cloning

Additional sequences we propose to improve translational performance

(1) To increase the efficiency of translational initiation

• Kozak sequence

• Shine-Dalgarno Sequence

(2) To increase the efficiency of translational termination

• Stop codon(TAA)


Composite Parts

BBa_K2567001: express nicA and degrade nicotine

Use a promoter (BBa_J23119), ribosome binding site (BBa_B0034), nicA coding region (BBa_K2567000), terminator(BBa_B0015). This part can express nicA, and nicA can degrade nicotine.