We designed 58 parts to accomplish our goal this year. Among them, 31 parts are composite parts and the rest are basic parts. We shared characterization of some important parts. All parts that we used had been submitted to the iGEM Registry. Composite parts are closely related to the gene pathways we design. The basic parts contains most of the enzyme components we have characterized. Among all parts we uploaded this year, our favorite basic parts is BBa_K4800015, and the favorite composite part is BBa_K4800008. Moreover, if you want to get more information about our parts, we strongly recommend you to visit the specific part pages.
| Name | Type | Description | Designer | Length |
|---|---|---|---|---|
| BBa_K4800015 | Coding | GabT | Chenxi Ma | 1281 |
| BBa_K4800004 | Coding | MmCAR(Q302E) | Yao Yao | 3549 |
| BBa_K4800007 | Coding | YciA | Chenxi Ma | 465 |
| BBa_K4800017 | Coding | MmCAR | Yao Yao | 3549 |
| BBa_K4800020 | Coding | MAB3367 | Yao Yao | 3531 |
| BBa_K4800021 | Coding | MpCAR | Yao Yao | 3561 |
| BBa_K4800022 | Coding | MSMEG5739 | Yao Yao | 3621 |
| BBa_K4800023 | Coding | MAB4714 | Yao Yao | 3576 |
| BBa_K4800024 | Coding | Adh1 | Yao Yao | 780 |
| BBa_K4800025 | Coding | Hsd36 | Yao Yao | 762 |
| BBa_K4800026 | Coding | Adhp | Yao Yao | 1011 |
| BBa_K4800027 | Coding | YahK | Yao Yao | 1050 |
| BBa_K4800028 | Coding | PatA | Yifei Wu | 1443 |
| BBa_K4800029 | Coding | Spo3471 | Yifei Wu | 1455 |
| BBa_K4800030 | Coding | SAV2585 | Yifei Wu | 1890 |
| BBa_K4800031 | Coding | QLH | Yifei Wu | 1392 |
| BBa_K4800032 | Coding | SpuC | Yifei Wu | 1455 |
| BBa_K4800058 | Coding | MmCAR(T390E) | Yifei Wu | 3549 |
| BBa_K4800010 | DNA | ackA-pta Donor | Wenbin Zhao | 1007 |
| BBa_K4800011 | DNA | YcjQ Donor | Yifei Wu | 1635 |
| BBa_K4800012 | DNA | ackA-pta sgRNA | Wenbin Zhao | 81 |
| BBa_K4800013 | DNA | YcjQ sgRNA | Wenbin Zhao | 81 |
| BBa_K4800016 | DNA | sfp | Yao Yao | 675 |
| BBa_K4800000 | Regulatory | trc promoter | Wenbin Zhao | 30 |
| BBa_K4800056 | Regulatory | T7 promoter | Wenbin Zhao | 20 |
| BBa_K4800001 | Terminator | rrnB T1 terminator | Yifei Wu | 87 |
| BBa_K4800061 | Terminator | T7 terminator | Qiming Zou | 48 |
| Name | Type | Description | Designer | Length |
|---|---|---|---|---|
| BBa_K4800008 | Composite | Ptrc-sfp-MmCAR(Q302E)-SpyTag-EutM-SpyCatcher-YahK | Yifei Wu | 7077 |
| BBa_K4800003 | Composite | Ptrc-sfp-MmCAR-YahK | Yao Yao | 6179 |
| BBa_K4800005 | Composite | MmCAR(Q302E)-SpyTag | Jiali Wang | 3636 |
| BBa_K4800018 | Composite | Ptrc-sfp-MmCAR(Q302E)-YahK | Qiming Zou | 6160 |
| BBa_K4800019 | Composite | EutM-SpyCatcher | Yifei Wu | 729 |
| BBa_K4800033 | Composite | pRSFDuet-MAB3367 | Qiming Zou | 7996 |
| BBa_K4800034 | Composite | pRSFDuet-MpCAR | Qiming Zou | 8026 |
| BBa_K4800035 | Composite | pRSFDuet-MSMEG5739 | Yao Yao | 8086 |
| BBa_K4800036 | Composite | pRSFDuet-MAB4714 | Yao Yao | 8041 |
| BBa_K4800037 | Composite | pTrc99a-Adh1 | Yao Yao | 4929 |
| BBa_K4800038 | Composite | pTrc99a-Hsd36 | Chenxi Ma | 4932 |
| BBa_K4800039 | Composite | pTrc99a-Adhp | Chenxi Ma | 5181 |
| BBa_K4800040 | Composite | pTrc99a-YahK | Chenxi Ma | 5206 |
| BBa_K4800041 | Composite | pTrc99a-PatA | Chenxi Ma | 5615 |
| BBa_K4800042 | Composite | pTrc99a-Spo3471 | Chenxi Ma | 5630 |
| BBa_K4800043 | Composite | pTrc99a-SAV2585 | Chenxi Ma | 6065 |
| BBa_K4800044 | Composite | pTrc99a-QLH | Chenxi Ma | 5567 |
| BBa_K4800045 | Composite | pTrc99a-SpuC | Chenxi Ma | 5630 |
| BBa_K4800046 | Composite | pTrc99a-GabT | Chenxi Ma | 5456 |
| BBa_K4800047 | Composite | pTrc99a-YahK-GabT | Chenxi Ma | 6555 |
| BBa_K4800048 | Composite | pTrc99a-YahK-PatA | Chenxi Ma | 6714 |
| BBa_K4800049 | Composite | pTrc99a-YahK-Spo3471 | Chenxi Ma | 6729 |
| BBa_K4800050 | Composite | pTrc99a-YahK-SAV2585 | Chenxi Ma | 7164 |
| BBa_K4800051 | Composite | pTrc99a-YahK-SpuC | Qiming Zou | 6729 |
| BBa_K4800052 | Composite | pTrc99a-YahK-QLH | Qiming Zou | 6666 |
| BBa_K4800053 | Composite | pACYCDuet-sfp-MmCAR-YahK | Chenxi Ma | 9648 |
| BBa_K4800054 | Composite | pTrc99a-davB-davA-GabT | Chenxi Ma | 8068 |
| BBa_K4800055 | Composite | pRSFDuet-sfp-MmCAR-YahK | Yifei Wu | 9482 |
| BBa_K4800057 | Composite | pRSFDuet-sfp-MmCAR(T390E)-Yahk | Yifei Wu | 9456 |
| BBa_K4800059 | Composite | pRSFDuet-sfp-MmCAR(Q302E)-YahK | Yifei Wu | 9456 |
| BBa_K4800062 | Composite | pRSFDuet-sfp-MmCAR(Q302E)-SpyTag-EutM-SpyCatcher-YahK | Yifei Wu | 10373 |
GabT, 4-aminobutyrate-2-oxoglutarate transaminase from Escherichia coli K-12 is the initial enzyme of one of two 4-aminobutyrate (GABA) degradation pathways.
We design and constructed the plasmid pTrc99a-GabT were transferred into strain E. coli BL21 to determine its express. SDS-PAGE results confirmed that the molecular weight of GabT protein was correct.
Figure 1 SDS-PAGE analysis of GabT expression in E. coli BL21
GabT catalyzes 5-aminovalerate to glutarate semialdehyde. As the instability and detection difficulty of glutarate semialdehyde, we determined the activity of GabT in a cascade reaction with aldehyde reductase of Yahk by detecting the 5-hydroxyvalerate production. The cell that contained empty plasmid was used as the control. As the results shown in Figure 2, GabT from E. coli exhibited the ammonia transfer activity towards 5-aminovalerate, and the product of 5-hydroxyvalerate was successfully detected.
Figure 2 Determining the activity of GabT by a cascade reaction
BBa_K4800008We adder a trc promoter on the basis of the original part. Moreover, the amino acid of MmCAR at 302nd position was mutated from glutamine to glutamic acid. EutM scaffold was also used for improving the catalytic activity. 82.5 mM 1,5-PDO was produced by the cells that expressed this part, which is 1.89-fold higher than the wild type control.
Figure 3. Determining the activity of the optimized part by detecting 1,5-PDO production and 5-hydroxyvalerate consumption in comparison to the wild type part in a whole cell catalysis process