PMID- 23997340
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20130902
LR  - 20211021
IS  - 0046-8991 (Print)
IS  - 0973-7715 (Electronic)
IS  - 0046-8991 (Linking)
VI  - 52
IP  - 3
DP  - 2012 Sep
TI  - Characterization of DLP12 Prophage Membrane Associated Protein: HolinGFP.
PG  - 464-71
LID - 10.1007/s12088-012-0278-7 [doi]
AB  - Lysis cassette genes from phages determine the final lytic event of the host 
      cells. The lysis cassette genes are conserved in phages and prophages. The 
      membrane associated holin from DLP12 prophage, available as a GFP fusion 
      construct, was shown to be overexpressed, using confocal microscopy analysis, in 
      bacterial cells. The protein expression caused cell death in E. coli AG1 strain 
      suggesting the protein was functional. The His-tag HolinGFP protein was purified 
      using cobalt affinity column and was eluted in the presence of different 
      non-ionic detergents DDM (n-dodecyl-ss-d-maltoside), LDAO 
      (Lauryldimethylamine-oxide), OG (n-octyl beta-d-glucopyranoside) and C12E9 (dodecyl 
      nonaoxyethylene ether). HolinGFP existed predominantly as a dimer in LDAO in 
      Superdex S200 gel filtration chromatography. Circular dichroism and fluorescence 
      spectroscopy of the fluorescent HolinGFP in all four detergents (C12E9, DDM, 
      LDAO, and OG) confirmed the folded state. Both dithiobis succinimidyl propionate 
      and gluteraldehyde crosslinking revealed the existence of higher order oligomers 
      and dimers. HolinGFP has been functionally and biophysically characterised and is 
      being explored for crystallographic structure determination.
FAU - Srividhya, K V
AU  - Srividhya KV
AD  - Department of Genetic Engineering, School of Biotechnology, Madurai Kamaraj 
      University, Madurai, 625021 Tamilnadu India.
FAU - Krishnaswamy, S
AU  - Krishnaswamy S
LA  - eng
PT  - Journal Article
DEP - 20120628
PL  - India
TA  - Indian J Microbiol
JT  - Indian journal of microbiology
JID - 0374703
PMC - PMC3460124
OTO - NOTNLM
OT  - Crosslinking
OT  - DLP12
OT  - HolinGFP
OT  - Oligomerisation
OT  - Prophage
EDAT- 2013/09/03 06:00
MHDA- 2013/09/03 06:01
PMCR- 2013/09/01
CRDT- 2013/09/03 06:00
PHST- 2011/04/19 00:00 [received]
PHST- 2012/06/04 00:00 [accepted]
PHST- 2013/09/03 06:00 [entrez]
PHST- 2013/09/03 06:00 [pubmed]
PHST- 2013/09/03 06:01 [medline]
PHST- 2013/09/01 00:00 [pmc-release]
AID - 278 [pii]
AID - 10.1007/s12088-012-0278-7 [doi]
PST - ppublish
SO  - Indian J Microbiol. 2012 Sep;52(3):464-71. doi: 10.1007/s12088-012-0278-7. Epub 
      2012 Jun 28.