Name: generation of null mutants to elucidate the role of bacterial glycosyltransferases in bacterial motility
Uploaded: 2022-03-11T13:45:00
Description: Watch this Scientific Journal Video about Generation of Null Mutants to Elucidate the Role of Bacterial Glycosyltransferases in Bacterial Motility at JoVE.com

इस काम को राष्ट्रीय अनुसंधान परिषद कनाडा द्वारा समर्थित किया गया था, योजना Nacional de I + D (Ministerio de Economía y Competitividad, स्पेन) के लिए और Generalitat de Catalunya (Centre de Referència en Biotecnologia) के लिए।

प्रक्रिया का योजनाबद्ध निरूपण चित्र 1 में दिखाया गया है।
1. एयरोमोनास में फ्लैगेला ग्लाइकोसिलेशन द्वीप (एफजीआई) की जैव सूचना विज्ञान पहचान
<ol><li>Aeromonas जीनोम में PseAc बायोसि...

<ol>
	<li>Sch&#228;ffer, C., Messner, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Emerging+facets+of+prokaryotic+glycosylation.">Emerging facets of prokaryotic glycosylation.</a> FEMS Microbiology Review. 41 (1), 49-91 (2017).</li><li>De Maayer, P., Cowan, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flashy+flagella:+flagellin+modification+is+relatively+common+and+highly+versatile+among+the+Enterobacteriaceae.">Flashy flagella: flagellin modification is relatively common and highly versatile among the Enterobacteriaceae.</a> BMC Genomics. 17, 377 (2016).</li><li>Nothaft, H., Szymanski, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protein+glycosylation+in+bacteria:+sweeter+than+ever.">Protein glycosylation in bacteria: sweeter than ever.</a> Nature Review Microbiology. 8 (11), 765-778 (2010).</li><li>Benz, I., Schmidt, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Never+say+never+again:+protein+glycosylation+in+pathogenic+bacteria.">Never say never again: protein glycosylation in pathogenic bacteria.</a> Molecular Microbiology. 45 (2), 267-276 (2002).</li><li>Guerry, P., Szymanski, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Campylobacter+sugars+sticking+out.">Campylobacter sugars sticking out.</a> Trends in Microbiology. 16 (9), 428-435 (2008).</li><li>Hug, I., Feldman, M. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analogies+and+homologies+in+lipopolysaccharide+and+glycoprotein+biosynthesis+in+bacteria.">Analogies and homologies in lipopolysaccharide and glycoprotein biosynthesis in bacteria.</a> Glycobiology. 21 (2), 138-151 (2011).</li><li>Iwashkiw, J. A., Vozza, N. F., Kinsella, R. L., Feldman, M. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pour+some+sugar+on+it:+the+expanding+world+of+bacterial+protein+O-linked+glycosylation.">Pour some sugar on it: the expanding world of bacterial protein O-linked glycosylation.</a> Molecular Microbiology. 89 (1), 14-28 (2013).</li><li>Szymanski, C. M., Wren, B. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protein+glycosylation+in+bacterial+mucosal+pathogens.">Protein glycosylation in bacterial mucosal pathogens.</a> Nature Review Microbiology. 3, 225-237 (2005).</li><li>Tan, F. Y. Y., Tang, C. M., Exley, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sugar+coating:+bacterial+protein+glycosylation+and+host-microbe+interactions.">Sugar coating: bacterial protein glycosylation and host-microbe interactions.</a> Trends Biochemistry Sciences. 40 (7), 342-350 (2015).</li><li>Lairson, L. L., Henrissat, B., Davies, G. J., Withers, S. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glycosyltransferases:+structures,+functions,+and+mechanisms.">Glycosyltransferases: structures, functions, and mechanisms.</a> Annual Review Biochemistry. 77, 521-555 (2008).</li><li>Lombard, V., Golaconda Ramulu, H., Drula, E., Coutinho, P. M., Henrissat, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+carbohydrate-active+enzymes+database+(CAZy)+in+2013.">The carbohydrate-active enzymes database (CAZy) in 2013.</a> Nucleic Acids Research. 42, 490-495 (2014).</li><li>Weerapana, E., Imperiali, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Asparagine-linked+protein+glycosylation:+from+eukaryotic+to+prokaryotic+systems.">Asparagine-linked protein glycosylation: from eukaryotic to prokaryotic systems.</a> Glycobiology. 16 (6), 91 (2006).</li><li>Faridmoayer, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extreme+substrate+promiscuity+of+the+Neisseria+oligosaccharyl+transferase+involved+in+protein+O-glycosylation.">Extreme substrate promiscuity of the Neisseria oligosaccharyl transferase involved in protein O-glycosylation.</a> Journal of Biological Chemistry. 283 (50), 34596-34604 (2008).</li><li>Wacker, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Substrate+specificity+of+bacterial+oligosaccharyltransferase+suggests+a+common+transfer+mechanism+for+the+bacterial+and+eukaryotic+systems.">Substrate specificity of bacterial oligosaccharyltransferase suggests a common transfer mechanism for the bacterial and eukaryotic systems.</a> Proceedings of the National Academy of Sciences of the United States of America. 103 (18), 7088-7093 (2006).</li><li>Scott, A. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flagellar+glycosylation+in+Burkholderia+pseudomallei+and+Burkholderia+thailandensis.">Flagellar glycosylation in Burkholderia pseudomallei and Burkholderia thailandensis.</a> Journal of Bacteriology. 193 (14), 3577-3587 (2011).</li><li>Thibault, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+the+carbohydrate+moieties+and+glycosylation+motifs+in+Campylobacter+jejuni+flagellin.">Identification of the carbohydrate moieties and glycosylation motifs in Campylobacter jejuni flagellin.</a> Journal of Biological Chemistry. 276 (37), 34862-34870 (2001).</li><li>Schirm, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structural,+genetic+and+functional+characterization+of+the+flagellin+glycosylation+process+in+Helicobacter+pylori.">Structural, genetic and functional characterization of the flagellin glycosylation process in Helicobacter pylori.</a> Molecular Microbiology. 48 (6), 1579-1592 (2003).</li><li>McNally, D. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeted+metabolomics+analysis+of+Campylobacter+coli+VC167+reveals+legionaminic+acid+derivatives+as+novel+flagellar+glycans.">Targeted metabolomics analysis of Campylobacter coli VC167 reveals legionaminic acid derivatives as novel flagellar glycans.</a> Journal of Biological Chemistry. 282 (19), 14463-14475 (2007).</li><li>Logan, S. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+novel+carbohydrate+modifications+on+Campylobacter+jejuni+11168+flagellin+using+metabolomics-based+approaches.">Identification of novel carbohydrate modifications on Campylobacter jejuni 11168 flagellin using metabolomics-based approaches.</a> FEBS Journal. 276 (4), 1014-1023 (2009).</li><li>Wilhelms, M., Fulton, K. M., Twine, S. M., Tom&#225;s, J. M., Merino, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+glycosylation+of+polar+and+lateral+flagellins+in+Aeromonas+hydrophila+AH-3.">Differential glycosylation of polar and lateral flagellins in Aeromonas hydrophila AH-3.</a> Journal of Biological Chemistry. 287 (33), 27851-27862 (2012).</li><li>Canals, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-structural+flagella+genes+affecting+both+polar+and+lateral+flagella-mediated+motility+in+Aeromonas+hydrophila.">Non-structural flagella genes affecting both polar and lateral flagella-mediated motility in Aeromonas hydrophila.</a> Microbiology. 153, 1165-1175 (2007).</li><li>Howard, S. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Campylobacter+jejuni+glycosylation+island+important+in+cell+charge,+legionaminic+acid+biosynthesis,+and+colonization+of+chickens.">Campylobacter jejuni glycosylation island important in cell charge, legionaminic acid biosynthesis, and colonization of chickens.</a> Infection and Immunity. 77 (6), 2544-2556 (2009).</li><li>Verma, A., Arora, S. K., Kuravi, S. K., Ramphal, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Roles+of+specific+amino+acids+in+the+N-terminus+of+Pseudomonas+aeruginosa+flagellin+and+of+flagellin+glycosylation+in+the+innate+immune+response.">Roles of specific amino acids in the N-terminus of Pseudomonas aeruginosa flagellin and of flagellin glycosylation in the innate immune response.</a> Infection and Immunity. 73 (12), 8237-8246 (2005).</li><li>Lamy, B., Baron, S., Barraud, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aeromonas:+the+multifaceted+middleman+in+the+One+Health+world.">Aeromonas: the multifaceted middleman in the One Health world.</a> Current Opinion in Microbiology. 65, 24-32 (2022).</li><li>Gav&#237;n, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lateral+flagella+of+Aeromonas+species+are+essential+for+epithelial+cell+adherence+and+biofilm+formation.">Lateral flagella of Aeromonas species are essential for epithelial cell adherence and biofilm formation.</a> Molecular Microbiology. 43 (2), 383-397 (2002).</li><li>Altschul, F. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gapped+BLAST+and+PSI-BLAST:+a+new+generation+of+protein+database+search+programs.">Gapped BLAST and PSI-BLAST: a new generation of protein database search programs.</a> Nucleic Acids Research. 25 (17), 3389-3402 (1997).</li><li>Forn-Cun&#237;, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Polar+flagella+glycosylation+in+Aeromonas:+Genomic+characterization+and+Involvement+of+a+specific+glycosyltransferase+(Fgi-1)+in+heterogeneous+flagella+glycosylation.">Polar flagella glycosylation in Aeromonas: Genomic characterization and Involvement of a specific glycosyltransferase (Fgi-1) in heterogeneous flagella glycosylation.</a> Frontiers in Microbiology. 11, 595697 (2021).</li><li>Milton, D. L., O'Toole, R., Horstedt, P., Wolf-Watz, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flagellin+A+is+essential+for+the+virulence+of+Vibrio+anguillarum.">Flagellin A is essential for the virulence of Vibrio anguillarum.</a> Journal of Bacteriol.ogy. 178 (5), 1310-1319 (1996).</li><li>Kov&#225;cs, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+Pseudomonas+pyocyanea+by+the+oxidase+reaction.">Identification of Pseudomonas pyocyanea by the oxidase reaction.</a> Nature. 178 (4535), 703 (1956).</li><li>Fulton, K. M., Mendoza-Barber&#225;, E., Twine, S. M., Tom&#225;s, J. M., Merino, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Polar+glycosylated+and+lateral+non-glycosylated+flagella+from+Aeromonas+hydrophila+strain+AH-1+(Serotype+O11).">Polar glycosylated and lateral non-glycosylated flagella from Aeromonas hydrophila strain AH-1 (Serotype O11).</a> International Journal of Molecular Sciences. 16 (12), 28255-28269 (2015).</li><li>Khider, M., Willassen, N. P., Hansen, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+alternative+sigma+factor+RpoQ+regulates+colony+morphology,+biofilm+formation+and+motility+in+the+fish+pathogen+Aliivibrio+salmonicida.">The alternative sigma factor RpoQ regulates colony morphology, biofilm formation and motility in the fish pathogen Aliivibrio salmonicida.</a> BMC Microbiology. 18 (1), 116 (2018).</li><li>Pawar, S. V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+genetic+tools+to+tackle+c-di-GMP-dependent+signalling+in+Pseudomonas+aeruginosa.">Novel genetic tools to tackle c-di-GMP-dependent signalling in Pseudomonas aeruginosa.</a> Journal of Applied Microbiology. 120 (1), 205-217 (2016).</li><li>Vander Broek, W., Chalmers, K. J., Stevens, M. P., Stevens, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+proteomic+analysis+of+Burkholderia+pseuomallei+Bsa+Type+III+secretion+system+effectors+using+hypersecreting+mutants.">Quantitative proteomic analysis of Burkholderia pseuomallei Bsa Type III secretion system effectors using hypersecreting mutants.</a> The American Society for Biochemistry and Molecular Biology. 14 (4), 905-916 (2015).</li></ol>

इस विधि का महत्वपूर्ण प्रारंभिक चरण फ्लैगेला और कल्पित जीटी के ग्लाइकोसिलेशन में शामिल क्षेत्रों की पहचान है क्योंकि ये एंजाइम उच्च होमोलॉजी दिखाते हैं और कई प्रक्रियाओं में शामिल होते हैं। सार्वज?...

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>ABI PRISM Big Dye Terminator v. 3.1 Cycle Sequencing Ready Reaction Kit</td>
			<td>Applied Biosystems</td>
			<td>4337455</td>
			<td>Used for sequencing</td>
		</tr>
		<tr>
			<td>AccuPrime Taq DNA Polymerase, high fidelity</td>
			<td>Invitrogen</td>
			<td>12346-086</td>
			<td>Used for amplification of AB, CD and AD fragments</td>
		</tr>
		<tr>
			<td>Agarose</td>
			<td>Conda-Pronadise</td>
			<td>8008</td>
			<td>Used for DNA electrophoresis</td>
		</tr>
		<tr>
			<td>Alkaline phosphatase, calf intestinal (CIAP)</td>
			<td>Promega</td>
			<td>M1821</td>
			<td>Used to remove phosphate at the 5&#8217; end</td>
		</tr>
		<tr>
			<td>Bacto agar</td>
			<td>Becton Dickinson</td>
			<td>214010</td>
			<td>Use for motility analysis</td>
		</tr>
		<tr>
			<td>BamHI</td>
			<td>Promega</td>
			<td>R6021</td>
			<td>Used for endonuclease restriction</td>
		</tr>
		<tr>
			<td>BglII</td>
			<td>Promega</td>
			<td>R6081</td>
			<td>Used for endonuclease restriction</td>
		</tr>
		<tr>
			<td>BioDoc-It Imagin System</td>
			<td>UVP</td>
			<td></td>
			<td>Bio-imaging station used for DNA visualization</td>
		</tr>
		<tr>
			<td>Biotaq polymerase</td>
			<td>Bioline</td>
			<td>BIO-21040</td>
			<td>Used for colony screening</td>
		</tr>
		<tr>
			<td>Cesium chloride</td>
			<td>Applichem</td>
			<td>A1126,0100</td>
			<td>Used for flagella purification</td>
		</tr>
		<tr>
			<td>Chloramphenicol</td>
			<td>Applichem</td>
			<td>A1806,0025</td>
			<td>Used for triparental mating</td>
		</tr>
		<tr>
			<td>Cytiva illustra GFX PCR DNA and Gel Band Purification Kit</td>
			<td>Cytivia</td>
			<td>28-9034-71</td>
			<td>Used for purification of PCR amplicons and DNA fragments.</td>
		</tr>
		<tr>
			<td>EDTA</td>
			<td>Applichem</td>
			<td>131026.1211</td>
			<td>Used for DNA electrophoresis</td>
		</tr>
		<tr>
			<td>Electroporation cuvettes 2 mm gap</td>
			<td>VWR</td>
			<td>732-1133</td>
			<td>Used for transformation</td>
		</tr>
		<tr>
			<td>Ethidium bromide</td>
			<td>Applichem</td>
			<td>A1152,0025</td>
			<td>Use for DNA visualization</td>
		</tr>
		<tr>
			<td>HyperLadder 1 Kb marker</td>
			<td>Bioline</td>
			<td>BIO-33053</td>
			<td>DNA marker</td>
		</tr>
		<tr>
			<td>Invitrogen Easy-DNA gDNA Purification Kit</td>
			<td>Invitrogen</td>
			<td>10750204</td>
			<td>Used for bacterial chromosomal DNA purification</td>
		</tr>
		<tr>
			<td>Luria-Bertani (LB) Miller agar</td>
			<td>Condalab</td>
			<td>996</td>
			<td>Used for Escherichia coli culture</td>
		</tr>
		<tr>
			<td>Luria-Bertani (LB) Miller broth</td>
			<td>Condalab</td>
			<td>1551</td>
			<td>Used for Escherichia coli culture</td>
		</tr>
		<tr>
			<td>Nanodrop ND-1000</td>
			<td>NanoDrop Techonologies Inc</td>
			<td></td>
			<td>Spectrophotometer used for DNA quantification</td>
		</tr>
		<tr>
			<td>Rifampicin</td>
			<td>Applichem</td>
			<td>A2220,0005</td>
			<td>Used for triparental mating</td>
		</tr>
		<tr>
			<td>SOC Medium</td>
			<td>Invitrogen</td>
			<td>15544034</td>
			<td>Used for electroporation recovery</td>
		</tr>
		<tr>
			<td>Spectinomycin</td>
			<td>Applichem</td>
			<td>A3834,0005</td>
			<td>Used for triparental mating</td>
		</tr>
		<tr>
			<td>SW 41 Ti Swinging-Bucket Rotor</td>
			<td>Beckman</td>
			<td>331362</td>
			<td>Used for flagella purification</td>
		</tr>
		<tr>
			<td>T4 DNA ligase</td>
			<td>Invitrogen</td>
			<td>15224017</td>
			<td>Used for ligation reaction</td>
		</tr>
		<tr>
			<td>Trypticasein soy agar</td>
			<td>Condalab</td>
			<td>1068</td>
			<td>Used for Aeromonas grown</td>
		</tr>
		<tr>
			<td>Trypticasein soy broth</td>
			<td>Condalab</td>
			<td>1224</td>
			<td>Used for Aeromonas grown</td>
		</tr>
		<tr>
			<td>Tryptone</td>
			<td>Condalab</td>
			<td>1612</td>
			<td>Use for motility analysis</td>
		</tr>
		<tr>
			<td>Tris</td>
			<td>Applichem</td>
			<td>A2264,0500</td>
			<td>Used for DNA electrophoresis and flagella purification</td>
		</tr>
		<tr>
			<td>Triton X-100</td>
			<td>Applichem</td>
			<td>A4975,0100</td>
			<td>Used for bacterial lysis</td>
		</tr>
		<tr>
			<td>Ultra Clear tubes (14 mm x 89 mm)</td>
			<td>Beckman</td>
			<td>344059</td>
			<td>Used for flagella purification</td>
		</tr>
		<tr>
			<td>Veriti 96 well Thermal Cycler</td>
			<td>Applied Biosystems</td>
			<td></td>
			<td>Used for PCR reactions</td>
		</tr>
		<tr>
			<td>Zyppy Plasmid Miniprep II Kit</td>
			<td>Zymmo research</td>
			<td>D4020</td>
			<td>Used for isolation of plasmid DNA</td>
		</tr>
	</tbody>
</table>

generation of null mutants to elucidate the role of bacterial glycosyltransferases in bacterial motility

प्रोकैरियोट्स में ग्लाइकोसिलेशन का अध्ययन एक तेजी से बढ़ता हुआ क्षेत्र है। बैक्टीरिया अपनी सतह पर विभिन्न ग्लाइकोसिलेटेड संरचनाओं को आश्रय देते हैं जिनके ग्लाइकन एक तनाव-विशिष्ट बारकोड का गठन करते हैं। संबद्ध ग्लाइकन यूकेरियोट्स की तुलना में चीनी संरचना और संरचना में उच्च विविधता दिखाते हैं और बैक्टीरिया-मेजबान मान्यता प्रक्रियाओं और पर्यावरण के साथ बातचीत में महत्वपूर्ण हैं। रोगजनक बैक्टीरिया में, ग्लाइकोप्रोटीन संक्रामक प्रक्रिया के विभिन्न चरणों में शामिल किए गए हैं, और ग्लाइकन संशोधन ग्लाइकोप्रोटीन के विशिष्ट कार्यों में हस्तक्षेप कर सकते हैं। हालांकि, ग्लाइकन संरचना, संरचना और जैवसंश्लेषण मार्गों की समझ में की गई प्रगति के बावजूद, रोगजनकता या पर्यावरण के साथ बातचीत में ग्लाइकोप्रोटीन की भूमिका की समझ बहुत सीमित है। इसके अलावा, कुछ बैक्टीरिया में, प्रोटीन ग्लाइकोसिलेशन के लिए आवश्यक एंजाइमों को अन्य पॉलीसेकेराइड बायोसिंथेटिक मार्गों के साथ साझा किया जाता है, जैसे कि लिपोपॉलीसेकेराइड और कैप्सूल बायोसिंथेटिक मार्ग। ग्लाइकोसिलेशन के कार्यात्मक महत्व को ग्लाइकोसिलेशन प्रक्रिया में शामिल होने के लिए माना जाने वाले विशिष्ट जीनों के उत्परिवर्तन के माध्यम से कई बैक्टीरिया में स्पष्ट किया गया है और लक्ष्य ग्लाइकोप्रोटीन की अभिव्यक्ति और संशोधित ग्लाइकन पर इसके प्रभाव का अध्ययन किया गया है। मेसोफिलिक एयरोमोनास में एक एकल और ओ-ग्लाइकोसिलेटेड ध्रुवीय फ्लैगेलम होता है। फ्लैगेलर ग्लाइकन एरोमोनास उपभेदों के बीच कार्बोहाइड्रेट संरचना और श्रृंखला की लंबाई में विविधता दिखाते हैं। हालांकि, आज तक विश्लेषण किए गए सभी उपभेदों में एक pseudaminic एसिड व्युत्पन्न को जोड़ने वाली चीनी के रूप में दिखाया गया है जो सेरीन या थ्रेओनिन अवशेषों को संशोधित करता है। ध्रुवीय फ्लैगेला असेंबली के लिए pseudaminic एसिड व्युत्पन्न की आवश्यकता होती है, और इसके नुकसान का आसंजन, बायोफिल्म गठन और उपनिवेशीकरण पर प्रभाव पड़ता है। इस लेख में विस्तृत प्रोटोकॉल का वर्णन है कि कैसे नल उत्परिवर्ती के निर्माण का उपयोग जीन या जीनोम क्षेत्रों की भागीदारी को समझने के लिए किया जा सकता है जिसमें फ्लैगेलर ग्लाइकन के जैवसंश्लेषण में कल्पित ग्लाइकोसिलट्रांसफरेज़ होते हैं। इसमें शामिल ग्लाइकोसिलट्रांसफरेज़ के कार्य और ग्लाइकन की भूमिका को समझने की क्षमता शामिल है। यह ग्लाइकन की कमी वाले उत्परिवर्ती की तुलना जंगली प्रकार के तनाव से करके प्राप्त किया जाएगा।

यह पद्धति एरोमोनास के जीन या क्रोमोसोमल क्षेत्रों में शून्य उत्परिवर्ती उत्पन्न करने के लिए एक प्रभावी प्रणाली प्रदान करती है जो फ्लैगेला ग्लाइकोसिलेशन और फ्लैगेला फिलामेंट की भूमिका को प्रभाव?...

Watch this Scientific Journal Video about Generation of Null Mutants to Elucidate the Role of Bacterial Glycosyltransferases in Bacterial Motility at JoVE.com

Generation of Null Mutants to Elucidate the Role of Bacterial Glycosyltransferases in Bacterial Motility

यहां, हम विशिष्ट ग्लाइकोसिलट्रांसफरेज़ या ग्लाइकोसिलट्रांसफरेज़ वाले क्षेत्रों में एयरोमोनास के शून्य उत्परिवर्ती के निर्माण का वर्णन करते हैं, गतिशीलता assays, और फ्लैगेला शुद्धिकरण एक ग्लाइकन के जैवसंश्लेषण में उनके एन्कोडेड एंजाइमों की भागीदारी और कार्य को स्थापित करने के लिए किया जाता है, साथ ही साथ जीवाणु रोगजनन में इस ग्लाइकन की भूमिका भी।

बैक्टीरियल गतिशीलता में बैक्टीरियल ग्लाइकोसिलट्रांसफरेज़ की भूमिका को स्पष्ट करने के लिए नल उत्परिवर्ती की पीढ़ी

The study of glycosylation in prokaryotes is a rapidly growing area. Bacteria harbor different glycosylated structures on their surface whose glycans ...

Research

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Immunology and Infection

Using Luciferase to Image Bacterial Infections in Mice

चूहे में छवि बैक्टीरियल संक्रमणों के लिए luciferase का उपयोग

Imaging is a valuable technique that can be used to monitor biological processes.  In particular, the presence of cancer cells, stem cells, specific ...

Measuring Bacterial Load and Immune Responses in Mice Infected with Listeria monocytogenes

Measuring Bacterial Load and Immune Responses in Mice Infected with Listeria monocytogenes

से संक्रमित चूहों में बैक्टीरियल लोड और इम्यून प्रतिक्रियाएँ माप लिस्टेरिया monocytogenes</em

Listeria monocytogenes (Listeria) is a Gram-positive facultative intracellular pathogen1. Mouse studies typically employ intravenous injection of ...

Introducing Shear Stress in the Study of Bacterial Adhesion

बैक्टीरियल आसंजन के अध्ययन में कतरें तनाव का परिचय

During bacterial infections a sequence of interactions occur between the pathogen and its host. Bacterial adhesion to the host cell surface is often the ...

Saliva, Salivary Gland, and Hemolymph Collection from Ixodes scapularis Ticks

Saliva, Salivary Gland, and Hemolymph Collection from Ixodes scapularis Ticks

लार लार ग्रंथि, Ixodes Scapularis Ticks से और Hemolymph संग्रह

Ticks are found worldwide and afflict humans with many tick-borne illnesses. Ticks are vectors for pathogens that cause Lyme disease and tick-borne ...

Bioluminescent Bacterial Imaging In Vivo

Bioluminescent Bacterial Imaging In Vivo

Bioluminescent बैक्टीरियल इमेजिंग<em&gt; Vivo में</em

This video describes the use of whole body bioluminesce imaging (BLI) for the study of bacterial trafficking in live mice, with an emphasis on the use of ...

Using the Overlay Assay to Qualitatively Measure Bacterial Production of and Sensitivity to Pneumococcal Bacteriocins

ओवरले परख का उपयोग गुणात्मक न्यूमोकोकल बैक्टीरियोसिन्स को बैक्टीरियल के उत्पादन और संवेदनशीलता उपाय

Streptococcus pneumoniae colonizes the highly diverse polymicrobial community of the nasopharynx where it must compete with resident organisms.  We have ...

Generation and Multi-phenotypic High-content Screening of Coxiella burnetii Transposon Mutants

Generation and Multi-phenotypic High-content Screening of Coxiella burnetii Transposon Mutants

जनरेशन और मल्टी-प्ररूपी उच्च सामग्री स्क्रीनिंग<em&gt; Coxiella burnetii</em&gt; Transposon म्यूटेंट

Invasion and colonization of host cells by bacterial pathogens depend on the activity of a large number of prokaryotic proteins, defined as virulence ...

Methods to Inhibit Bacterial Pyomelanin Production and Determine the Corresponding Increase in Sensitivity to Oxidative Stress

तरीके बैक्टीरियल Pyomelanin उत्पादन रोकना और ऑक्सीडेटिव तनाव के प्रति संवेदनशीलता में इसी वृद्धि निर्धारित करने के लिए

Pyomelanin is an extracellular red-brown pigment produced by several bacterial and fungal species. This pigment is derived from the tyrosine catabolism ...

Visualization of Twitching Motility and Characterization of the Role of the PilG in Xylella fastidiosa

Visualization of Twitching Motility and Characterization of the Role of the PilG in Xylella fastidiosa

हिल गतिशीलता के दृश्य और की भूमिका की विशेषता<em&gt; PilG</emमें&gt;<em&gt; Xylella fastidiosa</em

Xylella fastidiosa is a Gram-negative non-flagellated bacterium that causes a number of economically important diseases of plants. The twitching motility ...

Assays for Studying the Role of Vitronectin in Bacterial Adhesion and Serum Resistance

बैक्टीरियल आसंजन और सीरम प्रतिरोध में Vitronectin की भूमिका का अध्ययन करने के लिए परख

Bacteria utilize complement regulators as a means of evading the host immune response. Here, we describe protocols for evaluating the role vitronectin ...