Abstract

The C and D proteins from bacteriophage fd and fl have been purified and characterized. Since the DNA sequence is known, the amino acid composition of these purified proteins indicated that they are coded for by the phage, the C protein being the product of Gene IX and the D protein specified by Gene VI. The molecular weights of the C and D proteins were calculated from the DNA sequences to be 3,650 and 12,350, respectively. These values are close to the molecular weights observed after polyacrylamide gel electrophoresis of the proteins in sodium dodecyl sulfate. Since the C and D proteins can be selectively labeled with radioactive cysteine and arginine, it was possible to estimate the number of the C and D protein molecules relative to the number of A protein molecules which had been accurately determined in previous studies. Based on these results, the average phage particle contains 5 A, 5 D, and 10 C protein molecules.

Highlights

  • These values are close to the molecular weights observed after polyacrylamide gel electrophoresis of the proteins in sodium dodecyl sulfate

  • Over the years there have been several reports of the presence of other low molecular weight proteins associated with the virion [1,5,6,7]

  • Simons et al [8],using radiochemical techniques, have identified two proteins inM13 which were associated with the phage particles and had apparent molecular weights of3,500 and 11,500, respectively

Read more

Summary

EXPERIMENTAL PROCEDURES

Until recently there was direct evidence for only two capsid proteins, the major coat (B) and a minor coat (A) protein in the fdamentous viruses (fd, M13, and fl)(for virus, see Refs. 1-4). Over the years there have been several reports of the presence of other low molecular weight proteins associated with the virion [1,5,6,7]. Simons et al [8],using radiochemical techniques, have identified two proteins inM13 which were associated with the phage particles and had apparent molecular weights of500 and 11,500, respectively. Studieson the morphogenesis of filamentous phages require the isolation of phage-specific proteins in chemically significant quantities. This will allow specific antibodies to be prepared for determination of the location and time of appearance of these minor proteins. Goldsmith and Konigsberg [6] and Woolford et al [7] used suchan approach to definitively establish the identity of the A proteinas the product of Gene I11and tolocate this proteinat one end of the fd and fl virions

Materials
Growth a n d Purification of Radioactive Phage
RESULTS
30 Bottom
F R A C TNI OUNM B E R
DISCUSSION
Full Text
Paper version not known

Talk to us

Join us for a 30 min session where you can share your feedback and ask us any queries you have

Schedule a call

Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.