Abstract

T7 phage particles have an icosahedral head, a short conical tail, six tail fibers, a head-tail connector, and an internal core (as well as 39,937 base pairs of double-stranded DNA). In hopes of generating a more symmetric particle for X-ray crystallography, tailless T7 phage mutants were created. These were examined in the STEM and also sent to Mario Cerritelli at NIH who discovered, and we verified, that the particles prefer to orient themselves with the connector down on the thin carbon substrate. These oriented particles are amenable to cryo-EM studies and he is analyzing the organization of the DNA in them. It also turned out that these tailless phage gave particularly accurate mass measurements in the STEM. There is now a series of tailless phage mutants containing full-length DNA, an 8% deletion of DNA, and a 16% deletion of DNA. These are being used in the STEM to refine precision mass mapping and are being analyzed by cryo-EM at NIH.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR001777-13
Application #
5223600
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
13
Fiscal Year
1996
Total Cost
Indirect Cost

  Publications

Hind, Geoffrey; Wall, Joseph S; Várkonyi, Zsuzsanna et al. (2014) Membrane crystals of plant light-harvesting complex II disassemble reversibly in light. Plant Cell Physiol 55:1296-303
Garofalo, Robert; Mustanski, Brian; Johnson, Amy et al. (2010) Exploring factors that underlie racial/ethnic disparities in HIV risk among young men who have sex with men. J Urban Health 87:318-323
Joshi, V N; Mitra, D; England, M D et al. (2010) Large Covalently Linked Fluorescent and Gold Nanoparticle Immunoprobes. Microsc Microanal 16:966-967
Ackerson, Christopher J; Powell, Richard D; Hainfeld, James F (2010) Site-specific biomolecule labeling with gold clusters. Methods Enzymol 481:195-230
Hu, Minghui; Qian, Luping; Brinas, Raymond P et al. (2008) Gold nanoparticle-protein arrays improve resolution for cryo-electron microscopy. J Struct Biol 161:83-91
Garofalo, Robert; Mustanski, Brian; Donenberg, Geri (2008) Parents know and parents matter;is it time to develop family-based HIV prevention programs for young men who have sex with men? J Adolesc Health 43:201-4
Brinas, Raymond P; Hu, Minghui; Qian, Luping et al. (2008) Gold nanoparticle size controlled by polymeric Au(I) thiolate precursor size. J Am Chem Soc 130:975-82
Hu, Minghui; Zhang, Yian-Biao; Qian, Luping et al. (2008) Three-dimensional structure of human chromatin accessibility complex hCHRAC by electron microscopy. J Struct Biol 164:263-9
Garofalo, Robert; Mustanski, Brian S; McKirnan, David J et al. (2007) Methamphetamine and young men who have sex with men: understanding patterns and correlates of use and the association with HIV-related sexual risk. Arch Pediatr Adolesc Med 161:591-6
Mustanski, Brian; Garofalo, Robert; Herrick, Amy et al. (2007) Psychosocial health problems increase risk for HIV among urban young men who have sex with men: preliminary evidence of a syndemic in need of attention. Ann Behav Med 34:37-45

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