Is there is any possibility of the formation of TDP-rhmanose sugar in E. coli BL21 (DE3) if the glucose-1-phosphate thymidylyltransferase is knockdown? I appreciate the experts opinion and references regarding this study.
And also would appreciate if is not the case, how can I completely stop the synthesis of this sugar in E. coli?
Hi
If I have understood your question, I think you need to check the background of you strain. if it is already recombinant E.coli you need to extract the plasmid and screen the E.coli for this enzyme activity following high throughput screening. This article may give you an idea about high throughput screening. I hope that is useful.
http://www.jbc.org/content/286/15/13235.full.pdf+html
Best regards!
Dear Farah fadwa ben belgacem,
Thank you for your answer and the article recommendation. But I wanted to know how can I completely stop the synthesis of TDP-L-rhamnose in E. coli BL21 (DE3)? Since rfbA is an intermediate enzyme for TDP-glucose synthesis which ultimately converts into TDP-L-rhamnose by three successive enzymatic reaction.
Logically while rfbA deletion recombinant strain of E. coli BL21 (DE3), TDP-L-rhamnose must be absent but I am curious to know if there are any alternative pathway reported for this sugar synthesis in the same host?
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