How many transcripts per cell

timer Asked: Apr 23rd, 2015

Question description

(1)The polysaccharide degrading enzyme,levan fructotransferase(MW=46kD),can bemadein ayield of4g/LinrecombinantEcoli, usingaplasmidwith aROP-repressedColE1 origin of replication and astrongpromoter (lac). Assume you useastrong RBS sothat you get the maximum translation initiation rate (let’susethe numberdiscussed inclass – distance between ribosomes is120 nt). In order togetsuch ahigh yield of proteinitwas necessarytofindconditionsthat give avery highdensitycell culture (OD600of100). Assumethefollowing:transcription is induced at stationaryphase(so allcellsin theculturearemakingprotein)andthe mRNA transcript hasanaveragelifetimeof6min.

·Howmanytranscriptsper cell arerequired tosynthesize thisquantity ofproteinfor1L ofculture?

In our first lecture, we said that thereare ~10^4 mRNA transcripts (encoding ~1000differentproteins)in arapidlygrowingEcoli cell.

·Howdoesyourcalculated numberoftranscripts producedbyalac promoter compareto thistypical mRNA copynumberinacell?

Useful information:

ROP repressedColE1–50 copies/cellOD600of1forEcoli =8x10^8 cells/mL

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