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of antibiotics that inhibit cell wall biosynthesis. In this screen devised by Eugene
Dulaney, growing bacteria were exposed to test substances under hypertonic conditions. Inhibitors of cell wall synthesis caused growing bacteria to form spheroplasts. This screen enabled the discovery of
55:. The name spheroplast stems from the fact that after the microbe's cell wall is digested, membrane tension causes the cell to acquire a characteristic spherical shape. Spheroplasts are osmotically fragile, and will
276:, elongated cells that lack internal cross-walls. After a period of time, the cell walls of the filaments are digested, and the bacteria collapse into very large spheres surrounded by just their
31:
Gram-negative bacteria attempting to grow and divide in the presence of peptidoglycan synthesis-inhibiting antibiotics (e.g. penicillin) fail to do so, and instead end up forming spheroplasts.
345:
which makes extraction of cellular proteins difficult. Enzymatic digestion of the cell wall with zymolyase, creating spheroplasts, renders the cells vulnerable to easy
423:
Silver, L.L (2011). "Rational
Approaches to Antibacterial Discovery: Pre-Genomic Directed and Phenotypic Screening". In Dougherty, T.; Pucci, M.J. (eds.).
903:"Molecular template for a voltage sensor in a novel K channel. I. Identification and functional characterization of KvLm, a voltage-gated K channel from
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369:(PEG). With this method, nearly 100% of the animal cells may take up the foreign DNA. Upon conducting experiments following a modified
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680:
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animal cells. Spheroplasts with recombinant DNA are introduced into the media containing animal cells and are fused by
956:"Molecular and electrophysiological characterization of a mechanosensitive channel expressed in the chloroplasts of
469:"Morphological and ultrastructural changes in bacterial cells as an indicator of antibacterial mechanism of action"
170:. EDTA acts as a permeabilizer by binding to divalent ions such as Ca and removing them from the outer membrane.
163:
646:
Tortora, G.; Funke, B.; Case, C. (2016). "Chapter 4, Functional anatomy of prokaryotic and eukaryotic cells".
17:
701:"Inositol trisphosphate-dependent and -independent Ca mobilization pathways at the vacuolar membrane of
1019:"Dynamic oligomeric conversions of the cytoplasmic RCK domains mediate MthK potassium channel activity"
306:
1209:
281:
167:
71:
66:
When used to describe Gram-negative bacteria, the term spheroplast refers to cells from which the
860:
Blount, P.; Sukharev, S.I.; Moe, P.C.; Kung, C. (1999). "Mechanosensitive channels of bacteria".
314:
158:
causes Gram-negative bacteria to form spheroplasts, but only if a membrane permeabilizer such as
48:
1078:"Patch-clamp and phenotypic analyses of a prokaryotic cyclic nucleotide-gated K channel using
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and other excitable cells. To prepare giant spheroplasts, bacteria are treated with a
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675:(2nd ed.). United States of America: John Wiley & Sons, Inc. p. 234.
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Proceedings of the
National Academy of Sciences of the United States of America
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Proceedings of the
National Academy of Sciences of the United States of America
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Proceedings of the
National Academy of Sciences of the United States of America
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a particular channel to amplify its effect and make it easier to characterize.
209:
135:
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giant spheroplast for drug screening with automated planar patch clamp system"
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spheroplast can be used as an ion-channel expression system comparable to the
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In addition to the above antibiotics, inhibitors of protein synthesis (e.g.
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directly upstream of peptidoglycan synthesis induce spheroplasts too (e.g.
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377:, it was determined that spheroplasts may be able to transform at 4.9x10.
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1174:"The effect of spheroplast formation on the transformation efficiency in
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or lysozyme. According to some definitions, the term is used to describe
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Martinac, B.; Buechner, M.; Delcour, A.H.; Adler, J.; Kung, C. (1987).
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39:(or sphaeroplast in British usage) is a microbial cell from which the
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Fundamental
Laboratory Approaches for Biochemistry and Biotechnology
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Kikuchi, K.; Sugiura, M.; Nishizawa-Harada, C.; Kimura, T. (2015).
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260:, which was originally designed for characterizing the behavior of
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155:
323:
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650:(12th ed.). United States of America: Pearson. p. 84.
570:. The American Heritage Dictionary of the English Language. 2019
91:
convert Gram-negative bacteria into spheroplasts. These include
327:
346:
338:
56:
52:
953:
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51:. According to other definitions, the term also encompasses
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Nakayama, Y.; Fujiu, K.; Sokabe, M.; Yoshimura, K. (2007).
589:
587:
585:
901:
Santos, J.S.; Lundby, A.; Zazueta, C.; Montal, M. (2006).
864:. Methods in Enzymology. Vol. 294. pp. 458–482.
146:) also cause Gram-negative bacteria to form spheroplasts.
900:
292:
of the ion channels embedded in it. It is also common to
1017:
Kuo, M. M.-C.; Baker, K. A.; Wong, L.; Choe, S. (2007).
582:
43:
has been almost completely removed, as by the action of
558:
556:
427:. United States of America: Springer. pp. 33–75.
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Kuo, M. M.-C.; Saimi, Y.; Kung, C.; Choe, S. (2007).
859:
466:
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Cushnie, T.P.; O’Driscoll, N.H.; Lamb, A.J. (2016).
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349:with detergents or rapid osmolar pressure changes.
284:membranes. The membranes can then be analyzed on a
179:can be converted to spheroplasts using the enzymes
317:ion channels and it has been shown that the giant
208:used a spheroplast screen as a primary method for
82:
670:
166:is used to ease the enzyme's passage through the
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305:spheroplasts has been used to study the native
252:can be used to study the function of bacterial
138:) and inhibitors of folic acid synthesis (e.g.
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671:Ninfa, A.J.; Ballou, D.P.; Benore, M. (2009).
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245:Specially prepared giant spheroplasts of
241:spheroplast patched with a glass pipette.
341:cells are normally protected by a thick
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1172:Liu, I.; Liu, M.; Shergill, K. (2006).
313:. It has been extended to study other
199:
14:
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357:Bacterial spheroplasts, with suitable
299:The technique of patch clamping giant
473:Cellular and Molecular Life Sciences
425:Antibiotic Discovery and Development
272:). This causes the bacteria to form
799:"Pressure-sensitive ion channel in
709:The Journal of Biological Chemistry
699:Calvert, C.M.; Sanders, D. (1995).
373:protocol using calcium chloride in
361:inserted into them, can be used to
24:
387:Bacterial morphological plasticity
164:ethylenediaminetetraacetate (EDTA)
25:
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1199:
1133:"Genetic transformation of yeast"
1131:Gietz, R.D.; Woods, R.A. (2001).
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1086:Journal of Biological Chemistry
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204:From the 1960s into the 1990s,
83:Antibiotic-induced spheroplasts
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595:"Protoplasts and spheroplasts"
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1:
911:Journal of General Physiology
870:10.1016/s0076-6879(99)94027-2
648:Microbiology: An Introduction
402:
333:
95:synthesis inhibitors such as
1143:(4): 816–820, 822–826, 828.
111:. Antibiotics that inhibit
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621:"Definition of spheroplast"
433:10.1007/978-1-4614-1400-1_2
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288:apparatus to determine the
256:through a technique called
150:Enzyme-induced spheroplasts
10:
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766:10.1016/j.btre.2015.04.007
309:(MscL, MscS, and MscM) of
485:10.1007/s00018-016-2302-2
307:mechanosensitive channels
1210:Medical Subject Headings
746:"The application of the
601:. Encyclopedia.com. 2016
315:heterologously expressed
1044:10.1073/pnas.0609085104
985:10.1073/pnas.0609996104
722:10.1074/jbc.270.13.7272
627:. Merriam-Webster. 2019
625:www.merriam-webster.com
1099:10.1074/jbc.M703618200
905:Listeria monocytogenes
828:10.1073/pnas.84.8.2297
541:. Dictionary.com. 2019
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70:component but not the
49:Gram-negative bacteria
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923:10.1085/jgp.200609572
754:Biotechnology Reports
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195:Uses and applications
78:Spheroplast formation
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599:www.encyclopedia.com
200:Antibiotic discovery
113:biochemical pathways
109:β-lactam antibiotics
59:if transferred to a
1092:(33): 24294–24301.
1035:2007PNAS..104.2151K
976:2007PNAS..104.5883N
862:Ion Channels Part C
819:1987PNAS...84.2297M
367:polyethylene glycol
121:phosphoenolpyruvate
539:www.dictionary.com
243:
33:
1150:10.2144/01304rv02
970:(14): 5883–5888.
879:978-0-12-182195-1
682:978-0-470-08766-4
657:978-0-321-92915-0
479:(23): 4471–4492.
442:978-1-4614-1400-1
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353:Transfection
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254:ion channels
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213:fosfomycin,
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140:trimethoprim
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117:fosmidomycin
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18:Spheroplasts
294:overexpress
286:patch clamp
278:cytoplasmic
258:patch clamp
223:carbapenems
219:thienamycin
160:lactoferrin
154:The enzyme
89:antibiotics
37:spheroplast
1082:as a host"
494:10059/2129
403:References
392:Protoplast
334:Cell lysis
270:cephalexin
173:The yeast
134:, several
105:lactivicin
101:vancomycin
97:fosfomycin
63:solution.
45:penicillin
760:: 17–23.
363:transfect
343:cell wall
290:phenotype
274:filaments
266:septation
210:discovery
185:chitinase
61:hypotonic
41:cell wall
1226:Bacteria
1220:Category
1191:: 81–85.
1159:11314265
1118:17588940
1063:17287352
1004:17389370
941:16908725
784:28626710
631:July 21,
605:July 21,
574:July 21,
545:July 21,
513:27392605
504:11108400
381:See also
250:bacteria
181:lyticase
156:lysozyme
107:and the
87:Various
1109:3521034
1054:1892972
1031:Bibcode
995:1851586
972:Bibcode
932:2151562
888:9916243
847:2436228
815:Bibcode
775:5466043
731:7706267
521:2065821
375:E. coli
371:Hanahan
324:Xenopus
319:E. coli
311:E. coli
302:E. coli
262:neurons
1212:(MeSH)
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328:oocyte
239:E.coli
53:yeasts
1181:(PDF)
1178:DH5α"
517:S2CID
347:lysis
339:Yeast
282:outer
1155:PMID
1114:PMID
1059:PMID
1000:PMID
937:PMID
884:PMID
874:ISBN
843:PMID
780:PMID
727:PMID
677:ISBN
652:ISBN
633:2019
607:2019
576:2019
547:2019
509:PMID
437:ISBN
280:and
187:and
57:lyse
1145:doi
1104:PMC
1094:doi
1090:282
1049:PMC
1039:doi
1027:104
990:PMC
980:doi
968:104
927:PMC
919:doi
915:128
866:doi
833:PMC
823:doi
770:PMC
762:doi
717:doi
713:270
499:PMC
489:hdl
481:doi
429:doi
237:An
162:or
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431::
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