Labeling bacteriophage

Pierce CL, Rees JC, Fernandez FM, Barr JR. Viable Staphylococcus aureus quantitation using N-15 metabolically labeled bacteriophage amplification coupled with a multiple reaction monitoring proteomic workflow. Mol Cell Proteomics. 2012 11 M111.012849. doi 10.1074/mcp. Mlll.012849-5. 

Yano, R. Nogami, T. A method and an apparatus for detecting live bacteria using fluorescence-labeled bacteriophage. Jpn. Kokai Tokkyo Koho JP 11318499,1999. 

Fig. 3. Structure and sequence of repeats present in the fibrous proteins discussed in this chapter. (A) The adenovirus triple -spiral. A single repeat of one of the chains is shown as a stick model colored by atom, the other two as a secondary structure cartoon in yellow and orange. Amino acids contributing to the hydrophobic core are labeled, as is the glycine in the turn. (B) Triple -spiral sequence repeats. Conserved hydrophobic residues are indicated by a hash sign, the conserved glycine or proline by an asterisk. (C) The T4-hber fold. A single repeat of one of the chains is shown as a stick model colored by atom, the other two as a secondary structure cartoon in yellow and orange. Several of the conserved amino acids are labeled. (D) Repeating sequences present in bacteriophage T4 fiber proteins (Cerritelli et al., 1996). Conserved amino acids are indicated by a small letter conserved hydrophobic residues by a hash sign, and conserved small amino acids by a dot.

Additional support for DNA as the bearer of genetic information came from studies by Hershey and Chase (1952) in the replication of DNA-containing bacteriophages. Using virus particles isotopically labeled in either the viral DNA or in the viral protein, it was shown that the viral DNA entered the host cell (Escherichia coli), whereas the viral protein did not. Later experiments demonstrated that viral DNA alone was infectious and led to the formation of mature infectious virus particles of the appropriate genotype. The DNA contained all the information required for the synthesis of progeny phage particles. 

Figure 7.11 CE-LIF separation and quantitation of fluorescein-labeled lambda PCR product. A 500 bp product was generated with titrated amounts of lambda bacteriophage template and primers, one of which was fluorescein-labeled. The fragment was analyzed by CE-LIF, and only the fiuor from the PCR product was detected. With increasing amounts of DNA template, peak height and area of the product also increased up to a point, whereupon the PCR plateaued. Note the increase in primer and primer-dimer peaks as template availability decreases. (Reproduced with permission from KJ Ulfelder, Applications Information Bulletin A-1774, 1994. Copyright Beckman Instruments, Inc.)...

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