Achievements

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We have successful results:

• We helped to generate the data for the Interlab study this year.
• We isolated two full-length genes - sfp and YerP from our Bacillus subtilis strain and successfully processed them into biobricks, and the sequencing results are positive.
• We ligated lmrA into pHT01, a dual host, expression vector for Bacillus subtilis, and the sequencing result is positive. We measured the growth curve for our strain and tested two different transformation protocols.
• We assembled our surfactin-producing operon by overlap extension PCR, and the sequencing results are positive.
• We constructed a mathematical model and applied various estimations and confirmed parameters to obtain useful predictions.
• We designed and constructed a sprinkler model that can be used to spray bacterial suspensions for bioremediation.
• We adapted a protocol to measure leaching efficiency of different eluents in the soil quantitatively.

But we also have unsuccessful results:

• We failed to ligate many of our biobricks into pSB1C3 at various times, which may be due to low ligation efficiency in our lab.
• We didn’t do much characterization for our parts.
• We didn’t have time to test for surfactin production by our engineered strain.
• We didn’t construct our designed integration vector for B. subtilis, although we had prepared the fragments for Gibson assembly.
• We had a difficult time of transforming B. subtilis in our lab, and we once contaminated the competent cells.

Future Plans

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• We will complete the construction of our integration vector and surfactin-producing operon.
• We will test for the efficiency of surfactin production by analytical methods.
• We will refine our protocol for testing leaching efficiency and test our strain’s ability to emulsify the oil.
• We will analyze the eluate to see if the microbiome degrades the hydrocarbons.
• We will deposit our strain in a real-world contamination zone and see if it works.
• We will continue to analyze possible interactions between lmrA and surfactin as its substrate; then we will find mutated variants that could export surfactin more efficiently.