Studier ZYM-5052 Medium Kit. 1L Kit.
Studier Auto Induction Media is based on Studier’s ZYM-5052 media1 and has been reformulated for ease of use. All of the ingredients are contained in 2 bottles instead of the usual 5. At the time of usage, aseptically add 60mL of Studier Salts Solution (Cat. No.3S2600 40mL included) to Studier ZYM-5052 Medium for Auto-Induction. (Cat. No.3S2200 940mL included). Media is stable for 2 months once the salt solution has been added. Note: this media contains glucose, lactose, and glycerol as carbon sources. No further additions are required.
Auto induction media works with all T7 regulated E. coli vectors in BL21 and derivative strains.
PRINCIPLE OF OPERATION :
The original auto-induction paper by Studier1 can be found at the following link: http://wolfson.huji.ac.il/expression/local/bacteria/studierAutoInduced.pdf
When glucose is present in the media the expression of the lactose permease is inhibited and lactose will not be transported into the cell. After approximately 12 hrs of growth the glucose will be consumed and the lactose permease will begin to express thereby transporting lactose into the cells.
In strains such as BL21 the T7 RNA polymerase has been inserted into the chromosome and is under control of the lacUV5 promoter and is therefore inducible with lactose. Genes cloned into vectors containing the T7 promoter (eg. pET 2 and Flexi®3 vectors) will not be expressed until all of the glucose in the media has been consumed. This is very useful for protein production and generally results in significantly higher protein yields than is obtain in conventional media.
GROWTH CONDITIONS FOR HIGH THROUGHPUT [View PDF] :
1. F.W. Studier (2005) Protein production by autoâinduction in highâdensityshaking cultures. Prot. Exp. Pur. 41, 207â234.
2. pET vectors are available from EMD Millipore http://www.emdmillipore.com/life-science-research/vector-table-novagen-pet-vector-table/c_HdSb.s1O77QAAAEhPqsLdcab
3. Flexi® vectors are available from Promega. http://www.promega.com/resources/vector-sequences/bacterial-expression-vectors
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