Bacterial Genetic Systems - download pdf or read online

By Jeffrey H. Miller

ISBN-10: 0121821056

ISBN-13: 9780121821050

This quantity collates in a single resource method for in vivo genetic engineering and for genetic research in a variety of micro organism. not just is Escherichia coli good coated, yet so are different rising bacterial platforms

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Example text

Whether irradiation can equalize the frequency of h transduction for different markers, as it does for P1 transduction, has not been tested. Although the frequency of marker transduction with k appears to be about 100-fold lower than it is with P1, this is probably not due to packaging differences between the two phages, but rather reflects inefficient injection of DNA into recipient cells by h particles matured in oitro after headful packaging. Although h infectious particles inject their DNA into cells with near unit efficiency, h particles matured in oitro inject DNA into cells with an efficiency no greater than 5%.

35 H. Schmieger, Mol. Gen. Genet. 109, 323 (1970). 36 H. Schmieger and H. Backhaus, Mol. Gen. Genet. 120, 181 (1973). 37 j.

We use here the transduction of the argH defect in E. coli strain ABl157 (his-, leu-, argH-, proA-, thr-) to illustrate the transduction assay. Obviously the P1 transducing lysates should be prepared on an argH ÷ donor strain. Add 2-20 /zl of the P1 lysate (107-108 infectious phage) to 50/zl of a fresh overnight culture of ABl157 in 10 mM MgSO4 (1 × 109 cells/ml) and incubate the mixture for 10 min at 38° to permit the phage to adsorb to the cells. Now add 1 ml of L broth containing 10 mM 28 Escherichia coli AND Salmonella typhimurium [2] sodium citrate and incubate the infected cells at 38 ° for 1 hr with vigorous aeration.

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Bacterial Genetic Systems by Jeffrey H. Miller


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