Abstract
The main purpose of this project is to make C. tyrobutyricum facultative anaerobic, so that it can function well under anaerobic condition in gastrointestinal tract as a probiotics and under aerobic condition in alkaline soil as soil-improving bacteria. This goal can be achieved by introducing the dps and aceE genes into C. tyrobutyricum. dps protein can enhance the viability of the bacteria in aerobic environment mainly by protecting DNA. PDH expressed by aceE can substitute the main metabolic enzyme PFOR in the bacteria which is inactivated in the presence of oxygen.
To realize the above aim, we successfully constructed 7 parts. BBa_K4407001 and BBa_K4407002 are basic parts for dps and aceE genes. BBa_K4407010 to BBa_K4407013 are devices responsible for expressing dps and aceE genes using Pthl or Plac promoter. BBa_K4407014 and BBa_K4407015 are devices for co-expressing dps and aceE genes with Pthl or Plac promoter.
Table Part list
| No. | Name | Type | Description | Length |
|---|---|---|---|---|
| 1 | BBa_K4407001 | coding | dps,DNA protection against oxidative damage | 645 bp |
| 2 | BBa_K4407002 | coding | aceE,Alternate metabolic pathway | 2264 bp |
| 3 | BBa_K4407010 | device | Pthl-dps,Expression of dps with Pthl promoter | 1312 bp |
| 4 | BBa_K4407011 | device | Pthl-aceE ,Expression of aceE with Pthl promoter | 3331 bp |
| 5 | BBa_K4407012 | device | Plac-dps,Expression of dps in response to lactose | 1905 bp |
| 6 | BBa_K4407013 | device | Plac-aceE,Expression of aceE in response to lactose | 3924 bp |
| 7 | BBa_K4407014 | device | Plac-dps-aceE,Coexpression of dps and aceE in response to lactose | 4575 bp |
| 8 | BBa_K4407015 | device | Pthl-dps-aceE,Coexpression of dps and aceE with Pthl promoter | 3982 bp |
| 9 | BBa_K4407016 | regulatory | vgb promoter with another FNR binding site inserted, microaerobic | 149 bp |
