Optimized Expression of bTGase and 3C Protease in E. Coli

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Constructing pBAD/3C and pBAD/3C/bTGase Expression Plasmids

Prompt

An illustrative image that shows the construction of a plasmid. In the first part, a visual depiction of the pBAD/3C expression plasmid being constructed by cloning the 3C protease gene from B14 rhinovirus into pBAD/HisA, under the control of the araBAD promoter, being introduced into the cloning connector site between Sac I and Eco RI restriction enzymes at 5' and 3' ends respectively. The second part showcases the construction of the pBAD/3C/bTGase expression plasmid. The visual representation shows the T7 promoter and T7 terminator from pET20b (+) plasmid being subcloned into the pBAD/3C plasmid within Pci I and Bsm BI restriction sites at 5' and 3' ends respectively. Finally, the image hints at the gene fragment of bTGase from Bacillus amyloliquefaciens DSM7 and the prodomain of Streptomyces caniferus being cloned into pBAD/3C under the control of the T7 promoter.

Original Prompt: 2.3 . Construcción de plásmido Los genes que codifican la TGasa y la proteasa 3C se sintetizaron químicamente y la secuencia del gen que codifica la bTGasa de Bacillus amyloliquefaciens y la proteasa 3C del rinovirus humano se optimizó con codones para la expresión de E. coli . 2.3.1 . Primer inserto: construcción del plásmido de expresión pBAD/3C El plásmido de expresión pBAD/3C se construyó clonando el gen de la proteasa 3C del rinovirus B14 en pBAD/HisA bajo el control del promotor araBAD, introducido en el sitio conector de clonación entre las enzimas de restricción Sac I y Eco RI. en los extremos 5′ y 3′, respectivamente. 2.3.2 . Segundo inserto: construcción del plásmido pBAD/3C/bTGase El plásmido de expresión pBAD/3C/bTGase se construyó subclonando el promotor T7 y el terminador T7 del plásmido pET20b (+) en el plásmido pBAD/3C dentro de los sitios de restricción Pci I y Bsm BI en los extremos 5' y 3' respectivamente. El fragmento del gen bTGasa de Bacillus amyloliquefaciens DSM7 y el prodominio de Streptomyces caniferus se clonó en pBAD/3C bajo el control del promotor T7. Se insertó un sitio de escisión y reconocimiento de proteasa 3C entre el prodominio y la bTGasa. Para optimizar la expresión de bTGase, se colocó un codón para el aminoácido lisina (AAA) poco después del codón inicial del ATG.

Model: Imagen 4

Created on 1/21/2024 Report

Updated on 8/11/2025

License: Free to use with a backlink to Easy-Peasy.AI

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