Name: transformation of probiotic yeast and their recovery from gastrointestinal immune tissues following oral gavage in mice
Uploaded: 2016-02-08T14:00:00
Description: Watch this Scientific Journal Video about Transformation of Probiotic Yeast and Their Recovery from Gastrointestinal Immune Tissues Following Oral Gavage in Mice at JoVE.com

लेखकों इम्यूनोलॉजी और टीकों और एक एनआईएच नई अन्वेषक पुरस्कार (1DP2AI112242-01) के लिए बच्चों के केंद्र ट्रेसी जे मेमने को सम्मानित माध्यम से धन स्वीकार करते हैं। लेखकों को भी rad1 एस के उदार योगदान के लिए धन्यवाद Natalya पी Degtyareva cerevisiae।

1. यूवी mutagenesis Auxotrophic खमीर उपभेदों उत्पन्न करने के लिए <ol><li> पराबैंगनी विकिरण खुराक की जरूरत निर्धारित करने के लिए अस्तित्व घटता उत्पन्न <ol><li> मानक प्रक्रियाओं के अनुसार 26 YPD (खमीर निकालने peptone डेक्सट्रोज) मी?...

<ol>
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R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Methods in Enzymology, Volume 149: Guide to Yeast Genetics and Molecular Biology. , (1991).</li><li>DiCarlo, J. E., Norville, J. E., Mali, P., Rios, X., Aach, J., Church, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genome+engineering+in+Saccharomyces+cerevisiae+using+CRISPR-Cas+systems.">Genome engineering in Saccharomyces cerevisiae using CRISPR-Cas systems.</a> Nucleic acids research. 41 (7), 4336-4343 (2013).</li><li>Sander, J. D., Joung, J. 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साथ में, प्रोटोकॉल के साथ साथ आंत के लिए heterologous चिकित्सीय प्रोटीन की डिलीवरी के लिए विकास और auxotrophic प्रोबायोटिक खमीर उपभेदों के परीक्षण के लिए आवश्यक चरण का वर्णन है। इस हेरफेर और पुनः संयोजक प्रोबायोटिक ?...

<table border="0" cellpadding="0" cellspacing="0" width="704">
	<tbody>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">SmartSpec 3000 Spectrophotometer</td>
			<td style="width:107px;">BioRad</td>
			<td style="width:107px;">170-2501</td>
			<td style="width:275px;">Example of spectrophotometer for determining cell concentration and OD600 of yeast cultures</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">New Brunswick Roller Drum</td>
			<td style="width:107px;">Eppendorf</td>
			<td style="width:107px;">M1053-4004</td>
			<td style="width:275px;">Example of roller drum for yeast culture incubation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">UV Stratalinker 2400</td>
			<td style="width:107px;">Stratagene</td>
			<td style="width:107px;">400075-03</td>
			<td style="width:275px;">Example stratalinker</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Stuart Colony Counter SC6PLUS</td>
			<td style="width:107px;">11983044</td>
			<td style="width:107px;">Fisher Scientific</td>
			<td style="width:275px;">Plate stand with magnification records colony count upon sensing pressure from pen</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Scienceware Colony Counter&#160;</td>
			<td style="width:107px;">F378620002</td>
			<td style="width:107px;">Bel-Art Scienceware</td>
			<td style="width:275px;">Hand held colony counter pen</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Replica plating device</td>
			<td style="width:107px;">Fisherbrand</td>
			<td>09-718-1</td>
			<td rowspan="2" style="width:275px;">Example of replica plating stand and pads&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Velveteen squares</td>
			<td style="width:107px;">Fisherbrand</td>
			<td>09-718-2</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">&#160;L shaped sterile cell spreaders&#160;</td>
			<td style="width:107px;">Fisherbrand</td>
			<td style="width:107px;">14665230</td>
			<td style="width:275px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Deoxyribonucleic acid, single stranded from salmon testes</td>
			<td style="width:107px;">Sigma-Aldrich</td>
			<td style="width:107px;">D7656-1ML</td>
			<td style="width:275px;">Example carrier DNA for yeast LiOAc transformation</td>
		</tr>
		<tr height="126">
			<td height="126" style="height:126px;width:216px;">Gavage needles</td>
			<td style="width:107px;">Braintree Scientific</td>
			<td style="width:107px;">N-PK&#160;002</td>
			<td style="width:275px;">For mice 15-20 g, the suggested needle is a 22 gauge (1.25 mm ball), 1 in long, straight reusable gavage needle. For mice weighing greater than 20 g, 20 gauge or larger straight or curved gavage needles may be used</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">1mL sterile slip-tip disposable tuberculin syringe&#160;</td>
			<td style="width:107px;">Becton Dickinson</td>
			<td style="width:107px;">BD 309659</td>
			<td style="width:275px;"></td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Blunt forceps such as Electron Microscopy Sciences 7&#34; (178 mm) serrated tip, broad grip forceps</td>
			<td style="width:107px;">Electron Microscopy Sciences</td>
			<td style="width:107px;">77937-28</td>
			<td style="width:275px;">Example of blunt forceps needed for dissection&#160;</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Straight and curved dissection scissors&#160;</td>
			<td style="width:107px;">Electron Microscopy Sciences&#160;</td>
			<td style="width:107px;">72966-02 and 72966-03</td>
			<td style="width:275px;">Examples of scissors needed for dissection&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">IMDM</td>
			<td style="width:107px;">Life technologies</td>
			<td style="width:107px;">12440053</td>
			<td style="width:275px;"></td>
		</tr>
	</tbody>
</table>

transformation of probiotic yeast and their recovery from gastrointestinal immune tissues following oral gavage in mice

Development of recombinant oral therapy would allow for more direct targeting of the mucosal immune system and improve the ability to combat gastrointestinal disorders. Adapting probiotic yeast in particular for this approach carries several advantages. These strains have not only the potential to synthesize a wide variety of complex heterologous proteins but are also capable of surviving and protecting those proteins during transit through the intestine. Critically, however, this approach requires expertise in many diverse laboratory techniques not typically used in tandem. Furthermore, although individual protocols for yeast transformation are well characterized for commonly used laboratory strains, emphasis is placed here on alternative approaches and the importance of optimizing transformation for less well characterized probiotic strains. Detailing these methods will help facilitate discussion as to the best approaches for testing probiotic yeast as oral drug delivery vehicles and indeed serve to advance the development of this novel strategy for gastrointestinal therapy.

एक अस्तित्व वक्र पराबैंगनी विकिरण के बाद की पीढ़ी पतला खमीर कोशिकाओं है कि इस तरह अलग कॉलोनी बनाने इकाइयों (CFU) के रूप में करने में सक्षम हैं के चढ़ाना की आवश्यकता है। प्रत्येक 500 μl नमूने के र?...

Watch this Scientific Journal Video about Transformation of Probiotic Yeast and Their Recovery from Gastrointestinal Immune Tissues Following Oral Gavage in Mice at JoVE.com

Transformation of Probiotic Yeast and Their Recovery from Gastrointestinal Immune Tissues Following Oral Gavage in Mice

यहाँ प्रस्तुत तकनीकों कि, विकसित करने के लिए बदलने, प्रशासन, और प्रोबायोटिक खमीर Saccharomyces boulardii की heterologous प्रोटीन अभिव्यक्ति का परीक्षण किया जा सकता है के लिए एक एकीकृत वर्णन है।

प्रोबायोटिक खमीर के परिवर्तन और जठरांत्र इम्यून ऊतकों से उनके वसूली चूहे में मौखिक Gavage के बाद

Development of recombinant oral therapy would allow for more direct targeting of the mucosal immune system and improve the ability to combat ...

Research

JoVE Journal

Immunology and Infection

Seven Steps to Stellate Cells

सात कदम कक्ष तारामय

Hepatic stellate cells are liver-resident  cells of star-like morphology and are located in the space of Disse between  liver sinusoidal endothelial cells ...

Oral Transmission of Listeria monocytogenes in Mice via Ingestion of Contaminated Food

Oral Transmission of Listeria monocytogenes in Mice via Ingestion of Contaminated Food

की मौखिक संचरण<em&gt; लिस्टेरिया monocytogenes</em&gt; दूषित भोजन की घूस के माध्यम से चूहों में

L. monocytogenes are facultative intracellular bacterial pathogens that cause food borne infections in humans. Very little is known about the ...

Induction of Murine Intestinal Inflammation by Adoptive Transfer of Effector CD4+CD45RBhigh T Cells into Immunodeficient Mice

Induction of Murine Intestinal Inflammation by Adoptive Transfer of Effector CD4+CD45RBhigh T Cells into Immunodeficient Mice

Effector के सीडी 4 की दत्तक हस्तांतरण द्वारा Murine आंतों में सूजन की प्रेरण<sup&gt; +</sup&gt; CD45RB<sup&gt; उच्च</sup&gt; Immunodeficient चूहों में टी कोशिकाओं

There are many different animal models available for studying the pathogenesis of human inflammatory bowel diseases (IBD), each with its own advantages ...

In Situ Detection of Bacteria within Paraffin-embedded Tissues Using a Digoxin-labeled DNA Probe Targeting 16S rRNA

In Situ Detection of Bacteria within Paraffin-embedded Tissues Using a Digoxin-labeled DNA Probe Targeting 16S rRNA

-16 RRNA लक्ष्य निर्धारण एक Digoxin लेबल डीएनए जांच का उपयोग आयल एम्बेडेड ऊतकों के भीतर जीवाणुओं की सीटू का पता लगाने में

The presence of bacteria within the pocket epithelium and underlying connective tissue in gingival biopsies from patients with periodontitis has been ...

Isolation, Characterization and Functional Examination of the Gingival Immune Cell Network

अलगाव, लक्षण और मसूड़ों का प्रतिरक्षा सेल नेटवर्क के कार्यात्मक परीक्षा

Immune cell networks in tissues play a vital role in mediating local immunity and maintaining tissue homeostasis, yet little is known of the resident ...

Induction of Experimental Autoimmune Encephalomyelitis in Mice and Evaluation of the Disease-dependent Distribution of Immune Cells in Various Tissues

चूहे और विभिन्न ऊतकों में प्रतिरक्षा कोशिकाओं की रोग-निर्भर वितरण के मूल्यांकन में प्रायोगिक ऑटोइम्यून Encephalomyelitis की प्रेरण

Multiple sclerosis is presumed to be an inflammatory autoimmune disease, which is characterized by lesion formation in the central nervous system (CNS) ...

Protective Efficacy and Pulmonary Immune Response Following Subcutaneous and Intranasal BCG Administration in Mice

सुरक्षा प्रभावकारिता और फेफड़े के प्रतिरक्षा प्रतिक्रिया चूहे में चमड़े के नीचे और Intranasal बीसीजी प्रशासन निम्नलिखित

Despite global coverage of intradermal BCG vaccination, tuberculosis remains one of the most prevalent infectious diseases in the world. Preclinical data ...

Subcellular Fractionation from Fresh and Frozen Gastrointestinal Specimens

ताजा और जमे हुए जठरांत्र नमूनों से उपसेलुलर अंश

The purpose of this protocol is to fractionate human intestinal tissue obtained by endoscopy into nuclear and cytoplasmic compartments for the ...

Probiotic Studies in Neonatal Mice Using Gavage

नवजात चूहों में प्रोबायोटिक अध्ययन Gavage का उपयोग

Adult mouse models have been widely used to understand the mechanism behind disease progression in humans. The applicability of studies done in adult ...

Quantification of three DNA Lesions by Mass Spectrometry and Assessment of Their Levels in Tissues of Mice Exposed to Ambient Fine Particulate Matter

मास स्पेक्ट्रोमेट्री द्वारा तीन डीएनए घावों का परिमाणीकरण और परिवेशी महीन पार्टिकुलेट मैटर के संपर्क में आने वाले चूहों के ऊतकों में उनके स्तरों का आकलन

DNA adducts and oxidized DNA bases are examples of DNA lesions that are useful biomarkers for the toxicity assessment of substances that are ...