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By Jeffrey H. Miller

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 lined, yet so are different rising bacterial platforms

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This approach has the potential of revealing new genes in a pathway. Also, if carefully done, it can indicate those genes whose products interact. However, in some cases, ts mutations appear to give suppressor mutations in genes coding for products in a pathway not directly related to the original gene. For instance, mutations in a gene required for protein secretion could be suppressed by mutations in a host of genes involved in protein synthesis) 7 Furthermore, it is important to do careful allele specificity studies to establish significant interactions.

Mutant colonies which export the ribonuclease will degrade the RNA in the surrounding medium, giving a "halo" of clearing around the colonies. Mutants which do not make the enzyme or fail to export it will have no halo. This approach has been extended by using a leaky outer membrane mutant in another genetic screening procedure. In these mutant cells, an unstable hybrid membrane protein composed of the Tsr protein and alkaline phosphatase leaks the alkaline phosphatase portion into the growth medium.

11, 197 (1985). 27 M. Feiss and D. A. Siegele, Virology 92, 190 (1979). 28 N. Steinberg and R. Weisberg, Nature (London) 2,56, 97 (1975). 30 Escherichia coli AND Salmonella typhimurium Chromosomal DNA I +A Lambdapackagingheadful ["" [2] IP, "~ prohead +~ protelnsaCCess°ry packaging Filled head ~ + DNase Trimmed head + tails and accessory proteins Lembda Transducing particle Fro. 2. Generationof h transducingparticles. The steps are describedin the text. generate infectious particles (Fig. 2). As predicted by this model, the DNA packaged into h transducing particles is of uniform size and approximately 105-109% that of h wild type, presumably a X headful.

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