OD750 Measurement

7.1-7.30

Table 1: Growth curve of cyanobacteria Cultured under room temperature and natural light conditions.

OD750 Measurement

7.1-7.30

Table 2: Growth curve of cyanobacteria Cultured in a light incubator.

7.17

DH5α amplification of cscB gene and cscB plasmid extraction.

7.23

Plasmid extraction of cscB and transformation into PCC7942.

8.1

Measurement of cscB plasmid concentration and sample submission for testing.

8.29

SN medium used for wild type, pre-transformation, and post-transformation.

We measured sucrose content from September 7 to 22, showing that transformed cyanobacteria had decreased intracellular sucrose and increased supernatant sucrose, indicating their ability to secrete sucrose.

7.9

Designed a thiobarbituric acid method for colorimetric detection of formic acid concentration, using a microplate reader to measure OD450 absorbance.

7.14

Design three electrosynthesis experiments with wild-typeS. oneidensis MR-1.

7.22

Measurement of the resistance of different powered devices.

7.16-7.23

We prepared S. oneidensis MR-1 competent cells and attempted electroporation. However, the electroporation was unsuccessful.

7.24

Through continuous learning and optimization of the procedure, we successfully achieved electroporation.

8.1

We designed and completed a fourth electrosynthesis experiment, which confirmed the successful expression of the designed gene plasmid and improved EEU efficiency.

8.3

We designed the 5th and 6th electrosynthesis experiments, which confirmed that the addition of 2-HNQ improved EEU efficiency.

8.4

We designed the 7th and 8th electrosynthesis experiments, which confirmed that the addition of riboflavin improved EEU efficiency.

8.6

S. oneidensis MR-1 Chelex100 DNA Extraction

8.21

We completed the 9th and 10th electrosynthesis experiments, which confirmed that the addition of riboflavin and rGO improved EEU efficiency.

7.3

Designed four media formulations: 2216E, 2216E + 1.5% NaCl, 2216 + 3% NaCl, 2216E + 4.5% NaCl for cultivating V.N.

7.16

Attempted to Culture Vibrio natriegens with sodium formate.

7.29

Cabp-Chbd plasmid extraction.
First electroporation

8.2

1.Four sets of Cabp-Chbd plasmid extraction：

8.2

Electroporated plasmid.
100ng plasmid added to 50uL competent V.N.

8.30

V.N.
electroporation.
100ng plasmid added to 50uL competent V.N.

9.3

Extraction of CA plasmid from E. coli.

9.3

V.N. electroporation.

9.5

V.N. electroporation. transformation failure.

9.7

V.N. electroporation. transformation failure.

9.8

V.N. electroporation. transformation failure.

9.16

V.N. electroporation. transformation failure.

8.26

1.Glycerol stock of E.coli Cabp-Chbd plated on resistant medium.
2. E.coli Cabp-Chbd preserved by streaking.
3. Induced expression, fluorescence microscopy showed no 580nm excitation light. No fluorescence at 550nm. Reinduced and wrapped in aluminum foil, attempted fluorescence detection again with 535B microplate reader.

8.27

Preparation of induction expression system (4 sets).

9.5

0:00: Inoculated E. coli (containing Cabp-Chbd and CA genes), OD600 measured.

9.5

Measured mCherry and EGFP fluorescence intensity (morning)

9.5

Afternoon
significant change in fluorescence intensity, first induction failed.

9.16

Inoculated V.N., OD600 measured.

9.27

A concentration gradient of IPTG was prepared, and transformed Vibrio natriegens was inoculated at 22:00. When OD600 reached 0.5, IPTG was added to induce fluorescent protein expression, which was observed using a fluorescence microscope.

9.27

Upon excitation with red light under the fluorescence microscope, the Vibrio natriegens transformed with the mCherry gene exhibited a stable red fluorescent signal, confirming the successful expression of the plasmid.

9.30

Fluorescence microscopy of Vibrio natriegens expressing EGFP showed a strong, uniform green fluorescence under 488 nm excitation, with emission around 509 nm, confirming correct EGFP expression. This indicates successful transformation and effective transcription and translation of the CARPs plasmid in Vibrio natriegens.