barcode scanner code in c#.net VIRAL PHENOTYPES in Software

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Bacteriophage phenotypes fall generally into two categories: plaque morphology and growth characteristics on different bacterial strains. For example, T2, an E. coli phage (see g. 7.1), produces small plaques with fuzzy edges (genotype r ). Rapid-lysis mutants (genotype r) produce large, smooth-edged plaques ( g. 7.7). Similarly,
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Transformation was rst observed in 1928 by F. Grif th and was examined at the molecular level in 1944 by O. Avery and his colleagues, who used the process to demonstrate that DNA was the genetic material of bacteria. 9 presents the details of these experiments. In transformation, a cell takes up extraneous DNA found in the environment and incorporates it into its genome
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Figure 7.7 Normal (r ) and rapid-lysis (r) mutants of phage T2.
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Mottled plaques occur when r and r phages grow together. (From Molecular Biology of Bacterial Viruses by Gunther S. Stent. 1963,
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1978 by W. H. Freeman and Company. Used with permission.)
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Tamarin: Principles of Genetics, Seventh Edition
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II. Mendelism and the Chromosomal Theory
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7. Linkage and Mapping in Prokaryotes and Bacterial Viruses
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The McGraw Hill Companies, 2001
Sexual Processes in Bacteria and Bacteriophages
Bacterial sexual processes Exogenous DNA
E. coli DNA
Transformation
Conjugation F+ F
12.) Note that unlike eukaryotic crossing over, this is not a reciprocal process.The bacterial chromosome incorporates part of the foreign DNA. The remaining singlestranded DNA, originally part of the bacterial chromosome, is degraded by host enzymes called exonucleases; linear DNA is degraded rapidly in prokaryotes. Transformation is a very ef cient method of mapping in some bacteria, especially those that are inef cient in other mechanisms of DNA intake (such as transduction, discussed later in this chapter). For example, a good deal of the mapping of the soil bacterium, Bacillus subtilis, has been done through the process of transformation; E. coli, however, is inef cient in transformation, so other methods are used to map its chromosome.
Transformation Mapping
Hfr F
Phage
E. coli DNA from phage
Viral sexual process
Transduction
The general idea of transformation mapping is to add DNA from a bacterial strain with known genotype to another strain, also with known genotype, but with different alleles at two or more loci. We then look for incorporation of the donor alleles into the recipient strain of bacteria. The more often alleles from two loci are incorporated together into the host, the closer together these loci must be to each other. Thus, we can use an index of co-occurrence that is in inverse relationship to map distance: the larger the co-occurrence of alleles of two loci, the closer together the loci must be. This is another way of looking at the mapping concepts we discussed in
Viral recombination Transforming DNA Figure 7.8
Summary of bacterial and viral sexual processes.
(genetic material) through recombination. Not all bacteria are competent to be transformed, and not all extracellular DNA is competent to transform.To be competent to transform, the extracellular DNA must be doublestranded. To be competent to be transformed, a cell must have the surface protein, competence factor, which binds to the extracellular DNA in an energy-requiring reaction. However, bacteria that are not naturally competent can be treated to make them competent, usually by treatment with calcium chloride, which makes them more permeable.
Degraded
Bacterial chromosome
Cell division
Transformed cell
Mechanisms of Transformation
Under natural conditions, only one of the strands of extracellular DNA is brought into the cell. The single strand brought into the cell can then be incorporated into the host genome by two crossovers ( g. 7.9). (The molecular mechanisms of crossing over are presented in chapter
Figure 7.9 A single strand of transforming DNA (blue with a
allele) enters a bacterial cell (red chromosome with a allele). Two crossovers bring the foreign DNA into the bacterial chromosome. After DNA replication and cell division, one cell has the a allele and the other the a allele. The chromosome is drawn as a double circle, symbolizing the double-stranded structure of DNA.
Tamarin: Principles of Genetics, Seventh Edition
II. Mendelism and the Chromosomal Theory
7. Linkage and Mapping in Prokaryotes and Bacterial Viruses
The McGraw Hill Companies, 2001
Seven
Linkage and Mapping in Prokaryotes and Bacterial Viruses
chapter 6, where we discovered that the closer two loci are, the fewer the recombinations between them and thus the higher the co-occurrence. Now, we also must look at another concept, that is, selecting for recombinant cells. In fruit ies, every offspring of a mated pair represents a sampling of the meiotic tetrad, and thus a part of the total, whether or not recombination took place. Here, however, many cells are present that do not take part in the transformation process. In a bacterial culture, for example, only one cell in a thousand might be transformed.We must thus always be sure when working with bacterial gene transfer that we count only those cells that have taken part in the process. Let us look at an example. A recipient strain of B. subtilis is auxotrophic for the amino acids tyrosine (tyrA ) and cysteine (cysC ). We are interested in how close these loci are on the bacterial chromosome. We thus isolate DNA from a prototrophic strain of bacteria (tyrA cysC ). We add this donor DNA to the auxotrophic strain and allow time for transformation to take place ( g. 7.10). If the experiment is successful, and the loci are close enough together, then some of the recipient bacteria may incorporate donor DNA that has either both donor alleles or one or the
other donor allele. Thus, some of the recipient cells will now have the tyrA and cysC alleles, some will have just the donor tyrA allele, some will have just the donor cysC allele, and the overwhelming majority will be of the untransformed auxotrophic genotype, tyrA cysC . We thus need to count the transformed cells. We do this by removing any extraneous transforming DNA and then pouring the cells out onto a complete medium so that all cells can grow. These cells are then replica-plated onto three plates a minimal medium plate, a minimal medium plus tyrosine plate, and a minimal medium plus cysteine plate and allowed to grow overnight in an incubator at 37 C.We then count colonies ( g. 7.11).Those growing on minimal medium are of genotype tyrA cysC ; those growing on minimal medium with tyrosine but not growing on minimal medium are tyrA cysC ; and those growing on minimal medium with cysteine but not growing on minimal medium are tyrA cysC .The overwhelming majority will grow on complete medium, but not on minimal medium or minimal media with just tyrosine or cysteine added.This majority is made up of the nontransformants, that is, auxotrophs that were not involved in a transformation event they took up no foreign DNA.
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