PMID- 38082069
OWN - NLM
STAT- MEDLINE
DCOM- 20231219
LR  - 20231219
IS  - 1976-3794 (Electronic)
IS  - 1225-8873 (Linking)
VI  - 61
IP  - 11
DP  - 2023 Nov
TI  - A Method for Physical Analysis of Recombination Intermediates in Saccharomyces 
      cerevisiae.
PG  - 939-951
LID - 10.1007/s12275-023-00094-w [doi]
AB  - Meiosis is a process through which diploid cells divide into haploid cells, thus 
      promoting genetic diversity. This diversity arises from the formation of genetic 
      crossovers (COs) that repair DNA double-strand breaks (DSBs), through homologous 
      recombination (HR). Deficiencies in HR can lead to chromosomal abnormality 
      resulting from chromosomal nondisjunction, and genetic disorders. Therefore, 
      investigating the mechanisms underlying effective HR is crucial for reducing 
      genome instability. Budding yeast serves as an ideal model for studying HR 
      mechanisms due to its amenability to gene modifications and the ease of inducing 
      synchronized meiosis to yield four spores. During meiosis, at the DNA level, 
      programmed DSBs are repaired as COs or non-crossovers (NCOs) through structural 
      alterations in the nascent D-loop, involving single-end invasions (SEIs) and 
      double-Holliday junctions (dHJs). This repair occurs using homologous templates 
      rather than sister templates. This protocol, using Southern blotting, allows for 
      the analysis and monitoring of changes in DNA structures in the recombination 
      process. One-dimensional (1D) gel electrophoresis is employed to detect DSBs, 
      COs, and NCOs, while two-dimensional (2D) gel electrophoresis is utilized to 
      identify joint molecules (JMs). Therefore, physical analysis is considered the 
      most effective method for investigating the HR mechanism. Our protocol provides 
      more comprehensive information than previous reports by introducing conditions 
      for obtaining a greater number of cells from synchronized yeast and a method that 
      can analyze not only meiotic/mitotic recombination but also mitotic replication.
CI  - (c) 2023. The Author(s), under exclusive licence to Microbiological Society of 
      Korea.
FAU - Rhee, Kiwon
AU  - Rhee K
AD  - Department of Life Science, Chung-Ang University, Seoul, 06974, Republic of 
      Korea.
FAU - Choi, Hyungseok
AU  - Choi H
AD  - Department of Life Science, Chung-Ang University, Seoul, 06974, Republic of 
      Korea.
FAU - Kim, Keun P
AU  - Kim KP
AD  - Department of Life Science, Chung-Ang University, Seoul, 06974, Republic of 
      Korea. kpkim@cau.ac.kr.
FAU - Joo, Jeong H
AU  - Joo JH
AUID- ORCID: 0009-0006-0779-196X
AD  - Department of Life Science, Chung-Ang University, Seoul, 06974, Republic of 
      Korea. jkiiklovehot@naver.com.
LA  - eng
GR  - RS-2023-00208191/National Research Foundation of Korea/
GR  - 2022R1I1A1A01055532/National Research Foundation of Korea/
GR  - Research Scholarship Grants in 2022/Chung-Ang University/
PT  - Journal Article
DEP - 20231211
PL  - Korea (South)
TA  - J Microbiol
JT  - Journal of microbiology (Seoul, Korea)
JID - 9703165
RN  - 0 (Saccharomyces cerevisiae Proteins)
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - *Saccharomyces cerevisiae/genetics
MH  - *Saccharomyces cerevisiae Proteins/genetics
MH  - DNA Breaks, Double-Stranded
MH  - Meiosis
MH  - Homologous Recombination
MH  - DNA
OTO - NOTNLM
OT  - DNA physical analysis
OT  - Meiosis
OT  - Recombination
OT  - Saccharomyces cerevisiae
OT  - Southern blotting
EDAT- 2023/12/12 00:42
MHDA- 2023/12/19 06:41
CRDT- 2023/12/11 23:47
PHST- 2023/10/09 00:00 [received]
PHST- 2023/11/06 00:00 [accepted]
PHST- 2023/11/05 00:00 [revised]
PHST- 2023/12/19 06:41 [medline]
PHST- 2023/12/12 00:42 [pubmed]
PHST- 2023/12/11 23:47 [entrez]
AID - 10.1007/s12275-023-00094-w [pii]
AID - 10.1007/s12275-023-00094-w [doi]
PST - ppublish
SO  - J Microbiol. 2023 Nov;61(11):939-951. doi: 10.1007/s12275-023-00094-w. Epub 2023 
      Dec 11.