Metabolism of Amino Acids and Amines, Volume 17B: Part B by Nathan P. Kaplan, Nathan P. Colowick, Herbert Tabor, Celia

By Nathan P. Kaplan, Nathan P. Colowick, Herbert Tabor, Celia White Tabor

The severely acclaimed laboratory common, Methods in Enzymology, is among the such a lot hugely revered courses within the box of biochemistry. due to the fact 1955, every one quantity has been eagerly awaited, usually consulted, and praised via researchers and reviewers alike. The sequence includes a lot fabric nonetheless correct this present day - really a vital booklet for researchers in all fields of lifestyles sciences.

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In many pathways, blockage by mutation results in an accumulation of the intermediate preceding the block. These compounds can then be sought in the growth medium of this culture, and the accumulation of a particular compound (or of a particular chromatographically detectable substance) can serve as a means of classifying mutants. A more valuable variation of this, in terms of the ordering of genetic blocks in the pathway, is the use of cross feeding. When the excreted intermediate can he taken up by other cells, in some cases the compound accumulated by one mutant can serve as an alternate nutritional supplement for another.

43H. O. Smith and M. Levine, Virology 31,207 (1967). UES [1] 2. Preparation and Use of Phage PIkc with E. 01 M in sterile MgSO4 before use. Phage Preparation. Dilute full culture 43a 1:10 into LC-broth and grow for 1 hour. To 1 ml of culture add 107 phage particles and incubate 15 minutes for phage adsorption. 2 ml of mixture to 2 ml of melted LC soft agar and spread on an LC agar plate. Incubate plate at 37 ° overnight. Add 5 ml of Mg-tryptone broth to each plate and carefully emulsify the soft agar using a sterile glass rod.

If a genetic block of the pathway is selected in such a double regulatory mutant, the biosynthetic intermediate preceding the block should accumulate to very high levels. It may be possible in many pathways to use this method as a practical means of producing such compounds. The uses of amino acid analogs in the study of histidine biosynthesis can serve to illustrate applications of such compounds to genetic analysis of any pathway. Analogs of other amino acids and their effects have recently been reviewed.

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