Chemical mapping of DNA and counter-ion content inside phage by energy-filtered TEM
Nevsten, Pernilla LU ; Evilevitch, Alex LU
and Wallenberg, Reine
LU
(2012)
In Journal of Biological Physics
38(2).
p.229-240
- Abstract
- Double-stranded DNA in many bacterial viruses (phage) is strongly confined, which results in internal genome pressures of tens of atmospheres. This pressure is strongly dependent on local ion concentration and distribution within the viral capsid. Here, we have used electron energy loss spectroscopy (EELS), energy-filtered TEM (EFTEM) and X-ray energy dispersive spectroscopy to provide such chemical information from the capsid and the phage tail through which DNA is injected into the cell. To achieve this, we have developed a method to prepare thin monolayers of self-supporting virus/buffer films, suitable for EELS and EFTEM analysis. The method is based on entrapment of virus particles at air-liquid interfaces; thus, the commonly used... (More)
- Double-stranded DNA in many bacterial viruses (phage) is strongly confined, which results in internal genome pressures of tens of atmospheres. This pressure is strongly dependent on local ion concentration and distribution within the viral capsid. Here, we have used electron energy loss spectroscopy (EELS), energy-filtered TEM (EFTEM) and X-ray energy dispersive spectroscopy to provide such chemical information from the capsid and the phage tail through which DNA is injected into the cell. To achieve this, we have developed a method to prepare thin monolayers of self-supporting virus/buffer films, suitable for EELS and EFTEM analysis. The method is based on entrapment of virus particles at air-liquid interfaces; thus, the commonly used method of staining by heavy metal salts can be avoided, eliminating the risk for chemical artifacts. We found that Mg2 + concentration was approximately 2-4 times higher in the DNA-filled capsid than in the surrounding TM buffer (containing 10 mM Mg2 + ). Furthermore, we also analyzed the DNA content inside the phage tail by mapping phosphorus and magnesium. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/2571307
- author
- Nevsten, Pernilla
LU
; Evilevitch, Alex
LU
and Wallenberg, Reine
LU
- organization
- publishing date
- 2012
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Energy-Filtered Transmission Electron Microscopy (EFTEM), Electron, Energy Loss Spectroscopy (EELS), Magnesium, Phage lambda, Spermine, Chemical mapping
- in
- Journal of Biological Physics
- volume
- 38
- issue
- 2
- pages
- 229 - 240
- publisher
- Springer
- external identifiers
-
- wos:000302868400003
- scopus:84863859518
- pmid:23449697
- ISSN
- 0092-0606
- DOI
- 10.1007/s10867-011-9234-8
- language
- English
- LU publication?
- yes
- additional info
- The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Polymer and Materials Chemistry (LTH) (011001041), Biochemistry and Structural Biology (S) (000006142)
- id
- 7c4fc0a4-cc5d-462a-baff-1c31dcd0c7c5 (old id 2571307)
- date added to LUP
- 2016-04-01 13:06:19
- date last changed
- 2023-11-12 12:09:37
@article{7c4fc0a4-cc5d-462a-baff-1c31dcd0c7c5,
abstract = {{Double-stranded DNA in many bacterial viruses (phage) is strongly confined, which results in internal genome pressures of tens of atmospheres. This pressure is strongly dependent on local ion concentration and distribution within the viral capsid. Here, we have used electron energy loss spectroscopy (EELS), energy-filtered TEM (EFTEM) and X-ray energy dispersive spectroscopy to provide such chemical information from the capsid and the phage tail through which DNA is injected into the cell. To achieve this, we have developed a method to prepare thin monolayers of self-supporting virus/buffer films, suitable for EELS and EFTEM analysis. The method is based on entrapment of virus particles at air-liquid interfaces; thus, the commonly used method of staining by heavy metal salts can be avoided, eliminating the risk for chemical artifacts. We found that Mg2 + concentration was approximately 2-4 times higher in the DNA-filled capsid than in the surrounding TM buffer (containing 10 mM Mg2 + ). Furthermore, we also analyzed the DNA content inside the phage tail by mapping phosphorus and magnesium.}},
author = {{Nevsten, Pernilla and Evilevitch, Alex and Wallenberg, Reine}},
issn = {{0092-0606}},
keywords = {{Energy-Filtered Transmission Electron Microscopy (EFTEM); Electron; Energy Loss Spectroscopy (EELS); Magnesium; Phage lambda; Spermine; Chemical mapping}},
language = {{eng}},
number = {{2}},
pages = {{229--240}},
publisher = {{Springer}},
series = {{Journal of Biological Physics}},
title = {{Chemical mapping of DNA and counter-ion content inside phage by energy-filtered TEM}},
url = {{http://dx.doi.org/10.1007/s10867-011-9234-8}},
doi = {{10.1007/s10867-011-9234-8}},
volume = {{38}},
year = {{2012}},
}