Development and characterization of membrane surface display system using molecular chaperon, prsA, of Bacillus subtilis

2005 ◽  
Vol 334 (4) ◽  
pp. 1248-1253 ◽  
Author(s):  
June-Hyung Kim ◽  
In-Suk Park ◽  
Byung-Gee Kim
Keyword(s):  
Bacillus Subtilis ◽  
Membrane Surface ◽  
Surface Display ◽  
Display System ◽  
Molecular Chaperon

scholarly journals Novel Bacterial Membrane Surface Display System Using Cell Wall-Less L-Forms of Proteus mirabilis and Escherichia coli

2002 ◽  
Vol 68 (2) ◽  
pp. 525-531 ◽  
Author(s):  
Christian Hoischen ◽  
Christine Fritsche ◽  
Johannes Gumpert ◽  
Martin Westermann ◽  
Katleen Gura ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Cell Wall ◽  
Proteus Mirabilis ◽  
Fusion Proteins ◽  
Membrane Surface ◽  
Surface Display ◽  
Lactose Permease ◽  
Cell Fractionation ◽  
Display System ◽  
Reporter Protein

ABSTRACT We describe a novel membrane surface display system that allows the anchoring of foreign proteins in the cytoplasmic membrane (CM) of stable, cell wall-less L-form cells of Escherichia coli and Proteus mirabilis. The reporter protein, staphylokinase (Sak), was fused to transmembrane domains of integral membrane proteins from E. coli (lactose permease LacY, preprotein translocase SecY) and P. mirabilis (curved cell morphology protein CcmA). Both L-form strains overexpressed fusion proteins in amounts of 1 to 100 μg ml−1, with higher expression for those with homologous anchor motifs. Various experimental approaches, e.g., cell fractionation, Percoll gradient purification, and solubilization of the CM, demonstrated that the fusion proteins are tightly bound to the CM and do not form aggregates. Trypsin digestion, as well as electron microscopy of immunogold-labeled replicas, confirmed that the protein was localized on the outside surface. The displayed Sak showed functional activity, indicating correct folding. This membrane surface display system features endotoxin-poor organisms and can provide a novel platform for numerous applications.

scholarly journals Bacterial Surface Display of Levansucrase of Zymomonas mobilis Using Bacillus Subtilis Spore Display System

KSBB Journal ◽  
2011 ◽  
Vol 26 (3) ◽  
pp. 243-247
Author(s):  
June-Hyung Kim ◽  
Soo-Keun Choi ◽  
Heung-Chae Jung ◽  
Jae-Gu Pan ◽  
Byung-Gee Kim
Keyword(s):  
Bacillus Subtilis ◽  
Zymomonas Mobilis ◽  
Surface Display ◽  
Subtilis Spore ◽  
Display System ◽  
Bacterial Surface ◽  
Bacillus Subtilis Spore ◽  
Bacterial Surface Display

scholarly journals Surface Display of Recombinant Proteins on Bacillus subtilis Spores

2001 ◽  
Vol 183 (21) ◽  
pp. 6294-6301 ◽  
Author(s):  
Rachele Isticato ◽  
Giuseppina Cangiano ◽  
Hoa T. Tran ◽  
Annalisa Ciabattini ◽  
Donata Medaglini ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Surface Display ◽  
Surface Expression ◽  
Bioactive Molecules ◽  
Bacterial Spores ◽  
Fusion Partner ◽  
Display System ◽  
Dot Blot ◽  
System A ◽  
Powerful Approach

ABSTRACT We developed a novel surface display system based on the use of bacterial spores. A protein of the Bacillus subtilis spore coat, CotB, was found to be located on the spore surface and used as fusion partner to express the 459-amino-acid C-terminal fragment of the tetanus toxin (TTFC). Western, dot blot and fluorescent-activated cell sorting analyses were used to monitor TTFC surface expression on purified spores. We estimated that more than 1.5 × 103 TTFC molecules were exposed on the surface of each spore and recognized by TTFC-specific antibodies. The efficient surface presentation of the heterologous protein, together with the simple purification procedure and the high stability and safety record ofB. subtilis spores, makes this spore-based display system a potentially powerful approach for surface expression of bioactive molecules.

scholarly journals Development of a LytE-based high-density surface display system in Bacillus subtilis

2007 ◽  
Vol 1 (2) ◽  
pp. 177-190 ◽  
Author(s):  
Chyi-Liang Chen ◽  
Sau-Ching Wu ◽  
Wai Mui Tjia ◽  
Christopher L. C. Wang ◽  
Manfred J. Lohka ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Surface Display ◽  
High Density ◽  
Display System ◽  
Density Surface

Ultrastructural identification of three types of sensory setae on copepod antennae

1995 ◽  
Vol 53 ◽  
pp. 956-957
Author(s):  
T. M. Weatherby ◽  
P.H. Lenz
Keyword(s):  
Phase Locking ◽  
Membrane Surface ◽  
Reaction Times ◽  
High Sensitivity ◽  
Structural Features ◽  
Calanoid Copepods ◽  
Marine Food Webs ◽  
Dendritic Membrane ◽  
Ultrastructural Characterization

Crustaceans, as well as other arthropods, are covered with sensory setae and hairs, including mechanoand chemosensory sensillae with a ciliary origin. Calanoid copepods are small planktonic crustaceans forming a major link in marine food webs. In conjunction with behavioral and physiological studies of the antennae of calanoids, we undertook the ultrastructural characterization of sensory setae on the antennae of Pleuromamma xiphias.Distal mechanoreceptive setae exhibit exceptional behavioral and physiological performance characteristics: high sensitivity (<10 nm displacements), fast reaction times (<1 msec latency) and phase locking to high frequencies (1-2 kHz). Unusual structural features of the mechanoreceptors are likely to be related to their physiological sensitivity. These features include a large number (up to 3000) of microtubules in each sensory cell dendrite, arising from or anchored to electron dense rods associated with the ciliary basal body microtubule doublets. The microtubules are arranged in a regular array, with bridges between and within rows. These bundles of microtubules extend far into each mechanoreceptive seta and terminate in a staggered fashion along the dendritic membrane, contacting a large membrane surface area and providing a large potential site of mechanotransduction.

scholarly journals Sequence and characterization of Bacillus subtilis CheW.

1992 ◽  
Vol 267 (17) ◽  
pp. 12055-12060
Author(s):  
D.W. Hanlon ◽  
L.M. Márquez-Magaña ◽  
P.B. Carpenter ◽  
M.J. Chamberlin ◽  
G.W. Ordal
Keyword(s):  
Bacillus Subtilis
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