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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
PDF1- Construction of recombinant plasmid pMTL-Pthl-F/Xpk(BD)
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
PDF2-Construction of recombinant plasmid pMTL-Pthl-F/Xpk(QS)
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
PDF3-Construction of recombinant plasmids :pMTL-Pfba-F/Xpk、pMTL-Ptkt-F/Xpk
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
PDF4-Replacement of carbon sources and determination of butyric
acid
1. Using PUC57 vector plasmid as the template, F/Xpk(ASR) fragment (2436
bp) was amplifie;
2.Linearized vector pMTL-Pthl was obtained by PCR amplification using plasmid pMTL-Pthl-F/Xpk(BD)
as template;
3.The F/Xpk (ASR) gene fragment and the X-pMTL-Pthl linearized vector
were ligated by Gibson assembly.
Experimenter:Yijiu Lu,Pengfei Bai,Huaize Zhu
Recorder:Pengfei Bai
PDF5-Construction of recombinant plasmids :pMTL-Pthl-F_Xpk-ASR.pdf