PMID- 38009594
OWN - NLM
STAT- MEDLINE
DCOM- 20240214
LR  - 20240214
IS  - 1097-4644 (Electronic)
IS  - 0730-2312 (Linking)
VI  - 125
IP  - 2
DP  - 2024 Feb
TI  - The dyeing effect of acridine orange for multiple plasmid systems is sensitive to 
      temperature.
PG  - e30499
LID - 10.1002/jcb.30499 [doi]
AB  - The Goldview dyeing of the natural multiplasmid system of Lactobacillus plantarum 
      PC518 was affected by temperature. The article want to identify the specific 
      molecules that cause temperature sensitivity, then experiment on the universality 
      of temperature sensitivity, and finally preliminarily analyze the influencing 
      factors. At 5 degrees C and 25 degrees C, single pDNA, multiplasmid system, and linear DNA 
      samples were electrophoretic on agarose gel prestained by Goldview 1, 2, 3, and 
      acridine orange (AO), respectively. Eighteen vectors of Escherichia coli and two 
      vectors shortened by cloning were mixed into multiplasmid systems with different 
      member numbers, and then electrophoresis with AO staining was performed within 
      the range of 5 degrees C-45 degrees C, with a linearized multiplasmid system as the control. The 
      lane profiles (peaks) were captured with Image Lab 5.1 software. After 
      electrophoresis, the nine-plasmid-2 system was dyed with AO solutions of 
      different ionic strengths to detect the effect of ionic strength on temperature 
      sensitivity. It was measured that the UV-visible absorption spectra of 
      the nine-plasmid-2 system dissolved in AO solutions with different ionic 
      strengths and pH. Further, a response surface model was constructed using 
      Design-Expert.V8.0.6 software. The electrophoresis result showed that the 
      multiplasmid system from L. plantarum PC518 stained by AO staining showed a weak 
      band at 5 degrees C and five bands at 25 degrees C, which was similar to the result of staining 
      with Goldview 1, 2, and 3. The synthetic nine-plasmid-1 system and nine-plasmid-2 
      system displayed different band numbers on the electrophoresis gel in the 
      electrophoresis temperature range of 5 degrees C-45 degrees C, namely 3, 4, 6, 4, and 2 bands, as 
      well as 2, 6, 7, 8, and 5 bands. Using the 1x Tris-acetate-EDTA (TAE)-AO 
      solution, the poststaining results of the nine-plasmid-2 system in the 
      temperature range of 5 degrees C-45 degrees C were 4, 6, 9, 9, and 7 bands, respectively. 
      Further, using 5x, 10x, or 25x TAE buffer, the AO poststaining results at 5 degrees C 
      were 4, 2, and 1 bands, respectively. The ultraviolet spectral results from 5 degrees C 
      to 25 degrees C showed that there was a significant difference (3.5 times) in the 
      fluctuation amplitude at the absorption peak of 261.2 nm between 0x and 1-10x 
      TAE-AO solution containing the nine-plasmid-2 system. Specifically, the 
      fluctuation amplitudes of 0x, 1x, 5x, and 10x samples were 0.032, 0.109, 0.112, 
      and 0.110, respectively. At the same time, using 1x and 10x TAE buffer, the 
      AO-stained linear nine-plasmid-2 system remained stable and did not display 
      temperature sensitivity. The response surface models of the AO-stained 
      nine-plasmid-2 system intuitively displayed that the absorbance of the 1x TAE 
      samples increased significantly with increasing temperature compared to the 0x 
      TAE samples, regardless of the pH value. The findings confirmed a 
      temperature-dependent effect in AO staining of natural or synthetic multiplasmid 
      systems, with the optimum staining result occurring at 25 degrees C. Ion strength was a 
      necessary condition for the temperature sensitivity mechanism. This study layed 
      the groundwork for further investigation into the reasons or underlying 
      mechanisms of temperature sensitivity in AO staining of multiplasmid systems.
CI  - (c) 2023 Wiley Periodicals LLC.
FAU - Jiao, Qiuxia
AU  - Jiao Q
AD  - Department of Pathogenic Biology, Chengdu Medical College, Chengdu, China.
FAU - Zhang, Yumeng
AU  - Zhang Y
AD  - Department of Pathogenic Biology, Chengdu Medical College, Chengdu, China.
FAU - Xie, Juan
AU  - Xie J
AD  - Department of Pathogenic Biology, Chengdu Medical College, Chengdu, China.
FAU - Liu, Fang
AU  - Liu F
AD  - Department of Pathogenic Biology, Chengdu Medical College, Chengdu, China.
FAU - Peng, Chaoming
AU  - Peng C
AD  - Department of General Practice, The First Affiliated Hospital, Chengdu Medical 
      College, Chengdu, China.
FAU - Pan, Qu
AU  - Pan Q
AUID- ORCID: 0000-0003-3405-4544
AD  - Department of Pathogenic Biology, Chengdu Medical College, Chengdu, China.
LA  - eng
GR  - 21PJ112/Sichuan Provincial Health Commission Foundation/
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20231127
PL  - United States
TA  - J Cell Biochem
JT  - Journal of cellular biochemistry
JID - 8205768
RN  - F30N4O6XVV (Acridine Orange)
RN  - 0 (tris-acetate-EDTA buffer)
RN  - 0 (Coloring Agents)
RN  - 9G34HU7RV0 (Edetic Acid)
RN  - 0 (Acetates)
RN  - 0 (Ethylenediamines)
SB  - IM
MH  - *Acridine Orange/chemistry
MH  - *Coloring Agents
MH  - Temperature
MH  - Plasmids/genetics
MH  - Edetic Acid
MH  - *Acetates
MH  - *Ethylenediamines
OTO - NOTNLM
OT  - acridine orange
OT  - gel electrophoresis
OT  - multiple pasmid systems
OT  - temperature sensitivity
OT  - ultraviolet spectroscopy
EDAT- 2023/11/27 12:42
MHDA- 2024/02/12 15:44
CRDT- 2023/11/27 07:03
PHST- 2023/10/24 00:00 [revised]
PHST- 2023/05/14 00:00 [received]
PHST- 2023/11/06 00:00 [accepted]
PHST- 2024/02/12 15:44 [medline]
PHST- 2023/11/27 12:42 [pubmed]
PHST- 2023/11/27 07:03 [entrez]
AID - 10.1002/jcb.30499 [doi]
PST - ppublish
SO  - J Cell Biochem. 2024 Feb;125(2):e30499. doi: 10.1002/jcb.30499. Epub 2023 Nov 27.