By Kelly Thomas Hughes, Sidney P. Colowick, Nathan Oram Kaplan, Stanley R. Maloy
The significantly acclaimed laboratory average for greater than fifty years, tools in Enzymology is among the such a lot hugely revered courses within the box of biochemistry. when you consider that 1955, every one quantity has been eagerly awaited, usually consulted, and praised by way of researchers and reviewers alike. Now with over four hundred volumes (all of them nonetheless in print), the sequence includes a lot fabric nonetheless correct today-truly an important ebook for researchers in all fields of lifestyles sciences. This new quantity offers equipment concerning using bacterial genetics for genomic engineering. The e-book 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
It is possible to find transposon insertions in or near any gene of interest. 2. With very rare exceptions, transposon insertions result in complete loss of function. 3. The insertion mutation is completely linked to the phenotype of the transposon in genetic crosses. This makes it easy to transfer mutations into new strain backgrounds simply by selecting for transposon‐associated antibiotic resistance. METHODS IN ENZYMOLOGY, VOL. 421 Copyright 2007, Elsevier Inc. All rights reserved. 1016/S0076-6879(06)21002-4 transposons 12  4.
The EZ‐Tn5TM system is based on the hyperactive Tn5 system previously described by Goryshin and Reznikoff (1998). METHODS IN ENZYMOLOGY, VOL. 421 Copyright 2007, Elsevier Inc. All rights reserved. 1016/S0076-6879(06)21003-6 18 transposons  The EZ‐Tn5 Transposome contains the transposase (Tnp) covalently linked to the 19‐bp inverted repeat mosaic ends (ME) that define the transposon, and which flank a conditional origin of replication (R6K
ori) and a KanR kanamycin resistance gene. The Transposome is stable enough to be electroporated into any cell for which an electroporation protocol has been established.
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. 8. When used as a recipient in a genetic cross, transposon insertions behave as point mutations in fine‐structure genetic mapping. 9. Transposon insertions can be obtained that are near but not within a gene of interest.
Advanced Bacterial Genetics: Use of Transposons and Phage for Genomic Engineering by Kelly Thomas Hughes, Sidney P. Colowick, Nathan Oram Kaplan, Stanley R. Maloy