Genome editing in archaeal viruses and endogenous viral protein purification
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Genome editing in archaeal viruses and endogenous viral protein purification. / Alfastsen, Lauge; Peng, Xu; Bhoobalan-Chitty, Yuvaraj.
I: STAR Protocols, Bind 2, Nr. 4, 100791, 2021.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - Genome editing in archaeal viruses and endogenous viral protein purification
AU - Alfastsen, Lauge
AU - Peng, Xu
AU - Bhoobalan-Chitty, Yuvaraj
N1 - Publisher Copyright: © 2021 The Authors
PY - 2021
Y1 - 2021
N2 - Archaea-infecting viruses are morphologically and genomically among the most diverse entities. Unfortunately, they are also fairly understudied due to a lack of efficient genetic tools. Here, we present a detailed protocol for the CRISPR/Cas-based genome editing of the virus SIRV2 infecting the genus Sulfolobus, which could easily be adapted to other archaeal viruses. This protocol also includes the procedure for endogenous viral protein purification and identification, allowing for assessing the molecular mechanisms behind virus life cycle and virus-host interactions. For complete details on the use and execution of this protocol, please refer to Mayo-Muñoz et al. (2018) and Bhoobalan-Chitty et al. (2019).
AB - Archaea-infecting viruses are morphologically and genomically among the most diverse entities. Unfortunately, they are also fairly understudied due to a lack of efficient genetic tools. Here, we present a detailed protocol for the CRISPR/Cas-based genome editing of the virus SIRV2 infecting the genus Sulfolobus, which could easily be adapted to other archaeal viruses. This protocol also includes the procedure for endogenous viral protein purification and identification, allowing for assessing the molecular mechanisms behind virus life cycle and virus-host interactions. For complete details on the use and execution of this protocol, please refer to Mayo-Muñoz et al. (2018) and Bhoobalan-Chitty et al. (2019).
KW - CRISPR
KW - Genetics
KW - Microbiology
KW - Molecular Biology
KW - Protein Biochemistry
KW - Protein expression and purification
U2 - 10.1016/j.xpro.2021.100791
DO - 10.1016/j.xpro.2021.100791
M3 - Journal article
C2 - 34585154
AN - SCOPUS:85122827532
VL - 2
JO - STAR Protocols
JF - STAR Protocols
SN - 2666-1667
IS - 4
M1 - 100791
ER -
ID: 291362159