By Kelly Thomas Hughes, Sidney P. Colowick, Nathan Oram Kaplan, Stanley R. Maloy
The severely acclaimed laboratory typical for greater than fifty years, equipment in Enzymology is among the so much hugely revered courses within the box of biochemistry. on the grounds that 1955, each one quantity has been eagerly awaited, usually consulted, and praised through researchers and reviewers alike. Now with over four hundred volumes (all of them nonetheless in print), the sequence comprises a lot fabric nonetheless suitable today-truly a vital e-book for researchers in all fields of existence sciences. This new quantity provides equipment on the topic of using bacterial genetics for genomic engineering. The ebook contains sections on pressure collections and genetic nomenclature; transposons; and phage.
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Additional info for Advanced Bacterial Genetics: Use of Transposons and Phage for Genomic Engineering
Procedure 3. 2% D‐glucose and 15 g/l Bacto agar for solid media. LB (Luria broth): Per liter deionized water, 10 g tryptone, 5 g yeast extract, 5 g NaCl; add 15 g/l Bacto agar for solid medium. TTC‐indicator plates: Per liter deionized water, 10 g tryptone, 1 g yeast extract, 5 g NaCl, 15 g Bacto agar, 1% sugar, and 50 mg TTC. To prepare TTC‐ara indicator plates, add 10 g tryptone, 1 g yeast extract, 5 g NaCl, and 15 g Bacto agar to 500 ml of deionized water in a 1‐liter flask (Flask A). Add 10 g of L‐arabinose to 500 ml of deionized water in a second 1‐liter flask (Flask B).
116, 125–159. , and Gottesman, S. (1991). Uses of transposons with emphasis on Tn10. Methods Enzymol. 204, 139–180.  In Vivo Mutagenesis Using EZ‐Tn5TM By JOHN R. KIRBY Abstract Epicentre Biotechnologies has developed a suite of transposon‐based tools for use in modern bacterial genetics. This chapter highlights the EZ‐ Tn5TM TransposomeTM system and focuses on in vivo mutagenesis and subsequent rescue cloning. com/. Introduction The EZ‐Tn5TM TransposomeTM system from Epicentre provides a rapid and straightforward method for in vivo mutagenesis and target identification following rescue cloning from the desired mutant.
Transposon insertions in operons are nearly always strongly polar on expression of downstream genes. Thus, transposon insertions can be used to determine whether genes are in an operon. 6. Transposons can generate deletions nearby. This provides a convenient method for isolating adjacent deletion mutations. 7. Transposons can provide a portable region of homology for genetic recombination. Transposon insertions can be used to construct deletions or duplications with defined endpoints, or can serve as sites of integration of other genetic elements.
Advanced Bacterial Genetics: Use of Transposons and Phage for Genomic Engineering by Kelly Thomas Hughes, Sidney P. Colowick, Nathan Oram Kaplan, Stanley R. Maloy