Name: one-step negative chromatographic purification of helicobacter pylori neutrophil-activating protein overexpressed in escherichia coli in batch mode
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Description: Watch this Scientific Journal Video about One-step Negative Chromatographic Purification of Helicobacter pylori Neutrophil-activating Protein Overexpressed in Escherichia coli in Batch Mode at JoVE.co

We thank Dr. Chao-Sheng Cheng at National Tsing Hua University, Taiwan, for performing the circular dichroism measurement. We also thank Drs. Evanthia Galanis and Ianko D. Iankov at Mayo Clinic, USA, for providing the anti-HP-NAP monoclonal antibody. We appreciate Drs. Han-Wen Chang and Chung-Chu Chen at Mackay Memorial Hospital, Hsinchu, Taiwan, for providing advice for IRB application, Mr. Te-Lung Tsai at the Mackay Memorial Hospital, Hsinchu, Taiwan, for supervising the analysis of isolated neutrophils, and Ms. Ju-Chen Weng at National Tsing Hua University, Taiwan, for her technical assistance. This work was supported by grants from the Ministry of Science and Technology of Taiwan (MOST 104-2311-B-007-003, NSC101-2311-B-007-007 and NSC98-2311-B-007-006-MY3), the Joint Research Program of National Tsing Hua University and Mackay Memorial Hospital (100N7727E1, 101N2727E1, 103N2773E1), and the research program of National Tsing Hua University (104N2052E1).

Menneskelig blod ble samlet fra friske frivillige med skriftlig informert samtykke og godkjenning fra Institutional Review Board of National Tsing Hua University, Hsinchu, Taiwan. 1. Uttrykk av rekombinant HP-NAP i E. coli <ol><li> Forbered plasmid pET42a-NAP inneholder DNA-sekvensen av HP-NAP fra H. pylori 26695 belastning som tidligere beskrevet 16. Fremstille plasmider som inneholder DNA-sekvensen av HP-NAP med de ønskede punktmutasjoner som er beskrevet (se protokoll, trinn 8)...

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Den negative modus batch-kromatografi med DEAE anionbytteharpikser som presenteres her er egnet for rensing av rekombinant HP-NAP overuttrykkes i E. coli. PH-verdiene av bufferne som ble anvendt i trinnene med cellelysering og rensing er svært kritisk for å sikre løseligheten av HP-NAP i E. coli-lysater og effektiv separering av rekombinant HP-NAP fra vertscelleurenheter, henholdsvis. Bakterieceller skal lyseres ved pH 9,0, og den negative rensingen bør utføres ved pH 8,0 for å oppnå HP-NAP i h?...

Helicobacter pylori (H. pylori) er en viktig årsak til gastritt og magesår. Denne bakterien har også blitt klassifisert som kreftfremkallende hos mennesker ved International Agency for Research on Cancer, en del av Verdens helseorganisasjon, i 1994. Det har blitt anslått at forekomsten av H. pylori-infeksjon er 70% i utviklingsland og 30-40% i de industrialiserte landene 1. Selv om infeksjon rate av H. pylori er synkende i de industrialiserte landene, infeksjon rate på H. pylori i utviklingslandene er fortsatt høy 2. Standarden behandling for å utrydde H. pylori-infeksjon består av administrasjon av en protonpumpehemmer, PPI, og to antibiotika, klaritromycin pluss amoxicillin eller metronidazol tre. Men fremveksten av antibiotikaresistens hos H. pylori-relaterte sår terapi oppfordrer til utvikling av nye strategier for å forebygge or kurere infeksjonen. Utvikling av forebyggende og / eller terapeutisk vaksinasjon mot H. pylori kan tilveiebringe en alternativ tilnærming for å styre H. pylori-infeksjon. Helicobacter pylori nøytrofil-aktiverende protein (HP-NAP), en større virulens faktor av H pylori, ble først identifisert i vann ekstrakter av H. pylori med evne til å aktivere neutrofiler til å over til endotelceller og produsere reaktive oksygenarter (ROS) 4. Neutrofil infiltrasjon av mageslimhinnen som finnes i H. pylori- infiserte pasienter med aktiv gastritt kan føre til betennelse og vevskade av magen. Dermed kan HP-NAP spille en patologisk rolle ved å aktivere nøytrofile å indusere mage betennelse, noe som ytterligere forårsaker magesår eller H. pylori -associated mage sykdommer. Likevel er HP-NAP en potensiell kandidat for kliniske applikasjoner 5,6. På grunn av det immunogene og immunmodulerende proeiendommer av HP-NAP, kan dette proteinet brukes til å utvikle vaksiner, terapeutiske midler og diagnostiske verktøy. En klinisk studie er utført for ved hjelp av rekombinant HP-NAP som en av komponentene i et protein vaksine mot H. pylori. Denne vaksinen består av rekombinant HP-NAP, cytotoxin-assosiert gen A (CagA), og vacuolating cytotoxin A (Vaca) proteiner formulert med aluminiumhydroksid og har videre vist seg å være trygg og immunogen hos mennesker 7. Også HP-NAP fungerer som en potent immunomodulator å utløse T helper type 1 (Th1) -polarized immunresponser for kreftbehandling 8 og å nedregulere Th2-medierte immunresponser fremkalt av allergiske reaksjoner og parasittinfeksjoner 9,10. Som for diagnostikk, har rekombinant HP-NAP-baserte ELISA blitt brukt til å påvise antistoffer mot HP-NAP i H. pylori -infected pasienter 11. En studie viste at nivået av HP-NAP-spesifikke antistoffer i sera fra H. pylori -infected pasienter med magekreft var signifikant høyere enn den fra pasienter med kronisk gastritt 12. En annen studie viste også at serum antistoffer mot HP-NAP er forbundet med tilstedeværelse av ikke-cardia gastrisk adenokarsinom 13. Således kan rekombinant HP-NAP-baserte ELISA anvendes for å påvise antistoffer mot HP-NAP for prognose av magekreft i H. pylori -infected pasienter. Tatt sammen, kan det rensede HP-NAP videre benyttes for forebyggelse, behandling, og prognose av H. pylori -associated sykdommer samt kreftbehandling. Blant de mange metoder som benyttes for rensing av rekombinant HP-NAP uttrykt i Escherichia coli (E. coli) i sin native form rapportert hittil, et andre rensetrinn som omfatter gelfiltrering kromatografi er nødvendig for å oppnå høyrent HP-NAP 14-16 . Her, en metode under anvendelse av negativ modus satsvis kromatografi meddietylaminoetyl (DEAE) ionebytter-harpikser er beskrevet for rensing av HP-NAP overuttrykkes i E. coli med høy ytelse og høy renhet. Denne renseteknikken er basert på binding av vertscelleproteiner og / eller andre enn HP-NAP til harpiksen urenheter. Ved pH 8,0, er det nesten ingen andre proteiner unntatt HP-NAP utvinnes fra den ubundne fraksjonen. Denne rense tilnærming ved hjelp av DEAE ionebytterkromatografi i negativ modus er enkel og tidsbesparende ved å tillate rensing av rekombinant HP-NAP via ett-trinns kromatografi gjennom innsamling av den ikke-bundne fraksjon. I tillegg til HP-NAP, flere andre biomolekyler, for eksempel virus 17, immunoglobulin G (IgG) 18, hemoglobin 19, proteinfosfatase 20, og virulens faktor flagellin 21, har også blitt rapportert å bli renset ved ione-byttekromatografi på negativ modus. Den negative modus er foretrukket for ione-byttekromatografi hvis forurensninger er mindrekomponenter som er tilstede i prøven underkastet som skal renses 22. Anvendelsen av negative kromatografi i rensing av naturlige eller rekombinante biomolekyler har nylig blitt anmeldt 23. Denne rapporten gir en trinnvis protokoll for uttrykk av rekombinant HP-NAP i E. coli, lysis av cellene, og rensing av HP-NAP ved hjelp av negativ modus batch-kromatografi med DEAE-ionebytter-harpikser. Hvis et ønsket protein for rensning er egnet for ione-byttekromatografi i negativ modus, kan den beskrevne protokoll også være innrettet som et utgangspunkt for utviklingen av en renseprosess.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Material</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>pET42a-NAP&#160;</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>prepared as described in Supplementary data of Refernce 15&#160; 
			http://www.sciencedirect.com/science/article/pii/S0006291X08018317</td>
		</tr>
		<tr>
			<td>E. coli BL21 (DE3)</td>
			<td>Thermo Fisher Scientific Inc</td>
			<td>C6000-03</td>
			<td>https://www.thermofisher.com/order/catalog/product/C600003</td>
		</tr>
		<tr>
			<td>Kanamycin</td>
			<td>Amresco</td>
			<td>25389-94-0</td>
			<td>http://www.amresco-inc.com/KANAMYCIN-SULFATE-0408.cmsx</td>
		</tr>
		<tr>
			<td>Isopropyl &#946;-D-1-thiogalactopyranoside (IPTG)</td>
			<td>MD Biomedical Inc</td>
			<td>101-367-93-1</td>
			<td>http://www.antibody-antibodies.com/product_det.php?id=238064&#38;supplier=search&#38;name =IPTG%20</td>
		</tr>
		<tr>
			<td>phenylmethylsulfonyl fluoride (PMSF)</td>
			<td>Sigma-Aldrich</td>
			<td>10837091001</td>
			<td>protease inhibitor 
			http://www.sigmaaldrich.com/catalog/product/roche/PMSFRO?lang=en&#38;region=TW</td>
		</tr>
		<tr>
			<td>N-alpha-tosyl-L-lysinyl-chloromethylketone (TLCK)</td>
			<td>Sigma-Aldrich</td>
			<td>T7254</td>
			<td>protease inhibitor 
			http://www.sigmaaldrich.com/catalog/product/sigma/t7254?lang=en&#38;region=TW</td>
		</tr>
		<tr>
			<td>N-tosyl-L-phenylalaninyl-chloromethylketone (TPCK)</td>
			<td>Sigma-Aldrich</td>
			<td>T4376</td>
			<td>protease inhibitor 
			http://www.sigmaaldrich.com/catalog/product/sigma/t4376?lang=en&#38;region=TW</td>
		</tr>
		<tr>
			<td>Bio-Rad protein assay dye reagent concentrate</td>
			<td>Bio&#8208;Rad</td>
			<td>500-0006</td>
			<td>for protein quantitation 
			http://www.bio-rad.com/en-us/sku/5000006-bio-rad-protein-assay-dye-reagent-concentrate</td>
		</tr>
		<tr>
			<td>Protein standard (bovine serum albumin)</td>
			<td>Sigma-Aldrich</td>
			<td>P5619&#160;</td>
			<td>a standard protein for Bio-Rad Protein Assay 
			http://www.sigmaaldrich.com/catalog/product/fluka/p5619?lang=en&#38;region=TW</td>
		</tr>
		<tr>
			<td>DEAE&#8211;Sephadex &#160;A-25 chloride form</td>
			<td>Sigma-Aldrich</td>
			<td>A25120</td>
			<td>http://www.sigmaaldrich.com/catalog/product/sigma/a25120?lang=en&#38;region=TW</td>
		</tr>
		<tr>
			<td>Spectrum/Por dialysis tubing</td>
			<td>Spectrum Laboratories</td>
			<td>132720</td>
			<td>with molecular weight cutoff of 14 kDa 
			http://www.spectrumlabs.com/dialysis/RCtubing.html?Pn=132720;</td>
		</tr>
		<tr>
			<td>Acrodisc&#174; units with Mustang&#174; E membrane</td>
			<td>Pall</td>
			<td>MSTG25E3</td>
			<td>for endotoxin removal; operated at flow rates ranging from 1 to 4 ml/min 
			http://www.pall.com/main/laboratory/product.page?id=19992</td>
		</tr>
		<tr>
			<td>mouse monoclonal antibody MAb 16F4</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>raised against the purified HP-NAP of H. pylori strain NCTC 11637 as described in Refernce 23;&#160; A gift from Drs. Evanthia Galanis and Ianko D. Iankov at Mayo Clinic, USA 
			http://www.sciencedirect.com/science/article/pii/S0264410X10017585</td>
		</tr>
		<tr>
			<td>HiLoad 16/600 Superdex 200 pg</td>
			<td>GE Healthcare Life Sciences</td>
			<td>28989335</td>
			<td>for gel filtration chromatography 
			http://www.gelifesciences.com/webapp/wcs/stores/servlet/productById/en/GELifeSciences-tw/28989335</td>
		</tr>
		<tr>
			<td>PlusOne silver staining kit, protein</td>
			<td>GE Healthcare Life Sciences</td>
			<td>17-1150-01</td>
			<td>http://www.gelifesciences.com/webapp/wcs/stores/servlet/productById/en/GELifeSciences-tw/17115001</td>
		</tr>
		<tr>
			<td>Ficoll-Paque PLUS</td>
			<td>GE Healthcare Life Sciences</td>
			<td>17-1440-02</td>
			<td>for density gradient centrifugation to purify human neutrophils 
			http://www.gelifesciences.com/webapp/wcs/stores/servlet/catalog/en/GELifeSciences-tw/products/AlternativeProductStructure _16963/17144002</td>
		</tr>
		<tr>
			<td>Flat bottom 96-well white plate</td>
			<td>Thermo Fisher Scientific Inc</td>
			<td>236108</td>
			<td>http://www.thermoscientific.com/en/product/nunc-f96-microwell-black-white-polystyrene-plate.html</td>
		</tr>
		<tr>
			<td>Luminol</td>
			<td>Sigma-Aldrich</td>
			<td>A8511</td>
			<td>protected from light 
			http://www.sigmaaldrich.com/catalog/product/sigma/a8511?lang=en&#38;region=TW</td>
		</tr>
		<tr>
			<td>Expand &#160;long template PCR system</td>
			<td>Sigma-Aldrich</td>
			<td>11681834001</td>
			<td>source of High Fidelity PCR enzyme mix 
			http://www.sigmaaldrich.com/catalog/product/roche/elongro?lang=en&#38;region=TW</td>
		</tr>
		<tr>
			<td>Dpn I</td>
			<td>New England Biolabs</td>
			<td>R0176S</td>
			<td>https://www.neb.com/products/r0176-dpni</td>
		</tr>
		<tr>
			<td>Xho I</td>
			<td>New England Biolabs</td>
			<td>R0146S</td>
			<td>https://www.neb.com/products/r0146-xhoi</td>
		</tr>
		<tr>
			<td>E. coli DH5&#945;</td>
			<td>Thermo Fisher Scientific Inc</td>
			<td>18265-017</td>
			<td>https://www.thermofisher.com/order/catalog/product/18265017</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Product Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Equipment</td>
			<td></td>
			<td></td>
			<td> 
			&#160;</td>
		</tr>
		<tr>
			<td>U-2800 double beam UV/VIS spectrophotometer</td>
			<td>Hitachi&#160;</td>
			<td>N/A</td>
			<td>out of market and upgraded to a new model 
			http://hitachi-hta.com/products/life-sciences-chemical-analysis/uvvisible-spectrophotometers</td>
		</tr>
		<tr>
			<td>EmulsiFlex-C3 high pressure homogenizer</td>
			<td>Avestin Inc</td>
			<td>C315320</td>
			<td>http://www.avestin.com/English/c3page.html</td>
		</tr>
		<tr>
			<td>Hitachi Koki himac CP80WX general ultracentrifuge</td>
			<td>Hitachi Koki Co</td>
			<td>90106401</td>
			<td>for separation of the soluble and insoluble protein fractions from E. coli lysates 
			http://centrifuges.hitachi-koki.com/products/ultra/cp_wx/cp_wx.html</td>
		</tr>
		<tr>
			<td>&#196;KTA FPLC</td>
			<td>GE Healthcare Life Sciences</td>
			<td>18-1900-26</td>
			<td>for gel filtration chromatography 
			http://www.gelifesciences.com/webapp/wcs/stores/servlet/productById/en/GELifeSciences-tw/18190026</td>
		</tr>
		<tr>
			<td>Aviv model 62ADS CD spectrophotometer</td>
			<td>Aviv Biomedical</td>
			<td>N/A</td>
			<td>out of market and upgraded to a new model 
			http://www.avivbiomedical.com/circular.php</td>
		</tr>
		<tr>
			<td>LAS-3000 imaging system</td>
			<td>Fujifilm</td>
			<td>N/A</td>
			<td>discontinued and replaced 
			http://www.gelifesciences.com/webapp/wcs/stores/servlet/productById/en/GELifeSciences-tw/28955810</td>
		</tr>
		<tr>
			<td>Wallac 1420 (Victor2) multilabel counter</td>
			<td>Perkin-Elmer</td>
			<td>1420-018</td>
			<td>for chemiluminescence detection 
			http://www.perkinelmer.com/catalog/product/id/1420-018</td>
		</tr>
	</tbody>
</table>

one-step negative chromatographic purification of helicobacter pylori neutrophil-activating protein overexpressed in escherichia coli in batch mode

Helicobacter pylori nøytrofil-aktiverende protein (HP-NAP) er en viktig faktor virulens av Helicobacter pylori (H. pylori). Det spiller en avgjørende rolle i H. pylori-indusert gastrisk inflammasjon ved å aktivisere flere medfødte leukocytter, inkludert neutrofiler, monocytter og mastceller. De immunogene og immunmodulerende egenskaper av HP-NAP gjør det til en potensiell diagnostisk og vaksinekandidat for H. pylori og et nytt legemiddel kandidat for kreftbehandling. For å oppnå vesentlige mengder av renset HP-NAP som brukes for dens kliniske anvendelser, til en effektiv metode for å rense dette protein med høyt utbytte og renhet må etableres. I denne protokoll har vi beskrevet en fremgangsmåte for ett-trinns negativ kromatografisk rensing av rekombinant HP-NAP overuttrykt i Escherichia coli (E. coli) ved hjelp av dietylaminoetyl (DEAE) ionebytterharpikser (f.eks., Sephadex) in batch-modus. Rekombinant HP-NAP utgjør nesten 70% av det totale protein i E. coli og er nesten fullstendig gjenvunnet i den oppløselige fraksjonen ved cellelyse ved pH 9,0. Under optimale forhold ved pH 8,0, blir de fleste av HP-NAP gjenvunnet i den ubundne fraksjon, mens de endogene proteiner fra E. coli er effektivt fjernet av harpiksen. Denne rensemetode med negativt modus batch kromatografi med DEAE ionebytterharpikser gir funksjonell HP-NAP fra E. coli i sin opprinnelige form med høyt utbytte og renhet. Den rensede HP-NAP kan videre benyttes for forebyggelse, behandling, og prognose av H. pylori -associated sykdommer samt kreftbehandling.

Det skjematiske diagram av den eksperimentelle fremgangsmåten i negativ rensing av rekombinant HP-NAP uttrykt i E. coli ved hjelp av DEAE-ionebytter-harpikser i batch-modus, er vist i figur 1. Denne renseteknikken er basert på binding av vertscelleproteiner og / eller andre enn HP-NAP til harpiksen urenheter. Ved pH 8,0, er det nesten ingen andre proteiner unntatt HP-NAP i sin opprinnelige form utvinnes fra den ubundne fraksjonen (Figur 2A og ...

Watch this Scientific Journal Video about One-step Negative Chromatographic Purification of Helicobacter pylori Neutrophil-activating Protein Overexpressed in Escherichia coli in Batch Mode at JoVE.co

One-step Negative Chromatographic Purification of Helicobacter pylori Neutrophil-activating Protein Overexpressed in Escherichia coli in Batch Mode

Et høyt utbytte Fremgangsmåte for ett-trinns negativ rensing av rekombinant Helicobacter pylori nøytrofil-aktiverende protein (HP-NAP) overuttrykt i Escherichia coli ved hjelp av dietylaminoetyl harpikser i batch-modus, er beskrevet. HP-NAP renset ved denne fremgangsmåte er fordelaktig for utvikling av vaksiner, medikamenter, eller diagnostikk for H. pylori -associated sykdommer.

Ett-trinns Negativ kromatografiske rensing i<em&gt; Helicobacter pylori</em&gt; Neutrofilaktiverende Protein overuttrykkes i<em&gt; Escherichia coli</em&gt; I batch-modus

Helicobacter pylori neutrophil-activating protein (HP-NAP) is a major virulence factor of Helicobacter pylori (H. pylori). It plays a critical role in H. ...

Research

JoVE Journal

Immunology and Infection

One-step Negative Chromatographic Purification of Helicobacter pylori Neutrophil-activating Protein Overexpressed in Escherichia coli in Batch Mode

Rapid Identification of Gram Negative Bacteria from Blood Culture Broth Using MALDI-TOF Mass Spectrometry

Rapid Identifikasjon av gramnegative bakterier fra blodkultur buljong Bruke MALDI-TOF massespektrometri

An important role of the clinical microbiology laboratory is to provide rapid identification of bacteria causing bloodstream infection. Traditional ...

Non-Invasive Model of Neuropathogenic Escherichia coli Infection in the Neonatal Rat

Non-Invasive Model of Neuropathogenic Escherichia coli Infection in the Neonatal Rat

Non-invasiv Model of Neuropathogenic<em&gt; Escherichia coli</em&gt; Infeksjon i Neonatal Rat

Investigation of the interactions between animal host and bacterial pathogen is only meaningful if the infection model employed replicates the principal ...

High Resolution Electron Microscopy of the Helicobacter pylori Cag Type IV Secretion System Pili Produced in Varying Conditions of Iron Availability

High Resolution Electron Microscopy of the Helicobacter pylori Cag Type IV Secretion System Pili Produced in Varying Conditions of Iron Availability

High Resolution Elektronmikroskopi av<em&gt; Helicobacter pylori</em&gt; Cag Type IV Sekresjon System Pili Produsert i varierende forhold av Iron tilgjengelighet

Helicobacter pylori is a helical-shaped, gram negative bacterium that colonizes the human gastric niche of half of the human population1,2.  H. pylori is ...

Detection of Enterohemorrhagic Escherichia Coli Colonization in Murine Host by Non-invasive In Vivo Bioluminescence System

Detection of Enterohemorrhagic Escherichia Coli Colonization in Murine Host by Non-invasive In Vivo Bioluminescence System

Påvisning av enterohemorrhagiske Escherichia Coli kolonisering i Murine vert av ikke-invasiv i Vivo bioluminescens System

Enterohemorrhagic E. coli (EHEC) O157:H7, which is a foodborne pathogen that causesdiarrhea, hemorrhagic colitis (HS), and hemolytic uremic syndrome ...

Creation of a Dense Transposon Insertion Library Using Bacterial Conjugation in Enterobacterial Strains Such As Escherichia Coli or Shigella flexneri

Creation of a Dense Transposon Insertion Library Using Bacterial Conjugation in Enterobacterial Strains Such As Escherichia Coli or Shigella flexneri

Etableringen av et tett Transposon innsetting biblioteket bruker bakteriell bøyning i Enterobacterial stammer som Escherichia Coli eller Shigella flexneri

Transposon mutagenesis is a method that allows gene disruption via the random genomic insertion of a piece of DNA called a transposon. The protocol below ...

Overexpression and Purification of Human Cis-prenyltransferase in Escherichia coli

Overexpression and Purification of Human Cis-prenyltransferase in Escherichia coli

Overekspresjon og rensing av menneske<em&gt; Cis</em&gt; -prenyltransferase i<em&gt; Escherichia coli</em

Prenyltransferases (PT) are a group of enzymes that catalyze chain elongation of allylic diphosphate using isopentenyl diphosphate (IPP) via multiple ...

A Facile Protocol to Generate Site-Specifically Acetylated Proteins in Escherichia Coli

A Facile Protocol to Generate Site-Specifically Acetylated Proteins in Escherichia Coli

En lettvinte protokoll for å generere Site-Specifically Acetylated proteiner i Escherichia Coli

Post-translational modifications that occur at specific positions of proteins have been shown to play important roles in a variety of cellular processes. ...

Dissecting Multi-protein Signaling Complexes by Bimolecular Complementation Affinity Purification (BiCAP)

Dissecting Multi-protein Signaling Complexes by Bimolecular Complementation Affinity Purification BiCAP

Dissekere multi protein signalnettverk komplekser av resultatet av dette Complementation affinitet rensing (BiCAP)

The assembly of protein complexes is a central mechanism underlying the regulation of many cell signaling pathways. A major focus of biomedical research ...

Using Enhanced Green Fluorescence Protein-expressing Escherichia Coli to Assess Mouse Peritoneal Macrophage Phagocytosis

Using Enhanced Green Fluorescence Protein-expressing Escherichia Coli to Assess Mouse Peritoneal Macrophage Phagocytosis

Bruke forbedret grønne fluorescens Protein-uttrykke Escherichia Coli å vurdere musen Peritoneal Macrophage fagocytose

This manuscript describes a simple and reproducible method to perform a phagocytosis assay. The first part of this method involves building a ...

Purification of a High Molecular Mass Protein in Streptococcus mutans

Purification of a High Molecular Mass Protein in Streptococcus mutans

Rensing av en High Molecular Mass protein i Streptococcus mutans

Elucidation of a gene&#39;s function typically involves comparison of phenotypic traits of wild-type strains and strains in which the gene of interest has ...