2023_WSNJ-A

10/7-14/7 Construction of recombinant plasmid pMTL-Pthl-F/Xpk(BD)

1. PCR amplification on the genome of Clostridium acetobutylicum to obtain the fragment F/Xpk(BD).

2. Linearized vector pMTL-Pthl was obtained by PCR amplification using plasmid pMTL82151-Pthl-adhE2 as template;

3. pMTL-Pthl-F/Xpk(BD) was obtained by Gibson assembly

Experimenter：Pengfei Bai,Huaize Zhu,Ruige Li,Junjie Zheng,Youchang Zhang,Yuyao Cao,Yuntian Yang,Shufan Wu

Recorder：Huaize Zhu

1. The fragment F/Xpk(QS) was obtained by PCR amplification on Bifidobacterium adolescentis genome

2. Linearized vector pMTL-Pthl was obtained by PCR amplification using plasmid pMTL82151-Pthl-adhE2 as template;

3. pMTL-Pthl-F/Xpk(QS) was obtained by Gibson assembly

Experimenter：Pengfei Bai,Huaize Zhu,Junjie Zheng,Youchang Zhang,Yuyao Cao

Recorder：Huaize Zhu

1.PCR amplification on the genome of Clostridium tyrobutyricum to obtain the fragment Pfba、Ptkt.

2.Linearized vector pMTL-F/Xpk(BD) was obtained by PCR amplification using plasmid pMTL-Pthl-F/Xpk(BD) as template;

3.pMTL-Pfba-F/Xpk(BD)、pMTL-Ptkt-F/Xpk(BD) was obtained by Gibson assembly

Experimenter：Leyi Zhang,Yijiu Lu,Xiaoran Zhao,Yixuan Liu,Qingyue Feng,Ruoxi Liu,Jiale Li

Recorder：Leyi Zhang

1.Culture of Clostridium tyrobutyricum engineered bacteria Ct (Pthl F/Xpk-BD) using TGY medium supplemented with different carbon sources

2.Fermentation broths of Ct (Pthl F/Xpk-BD) in media containing different carbon sources were collected and assayed for product content using HPLC

3.Analysis of HPLC data

Experimenter：Yijiu Lu,Pengfei Bai,Huaize Zhu,Leyi Zhang，Shuyan Zhou,Keyu Chen,Zexi Zheng,Yixuan Liu

Recorder：Yijiu Lu