Name: a protocol to infect caenorhabditis elegans with salmonella typhimurium
Uploaded: 2014-06-26T15:00:00
Description: Watch this Scientific Journal Video about A Protocol to Infect Caenorhabditis elegans with Salmonella typhimurium at JoVE.com

我们感谢博士戴安娜Baronas洛厄尔的手稿的批判性阅读。这项工作得到了科学种子格兰特的FAU查尔斯·E·施密特学院，并从埃利森医学基金会的老化奖学金KJ支持

1，XLD（木糖赖氨酸去氧胆酸盐）琼脂平板 XLD琼脂是沙门氏菌，这似乎是在XLD琼脂平板上菌落黑色选择性生长培养基。然而，如果有污染的担忧，一个普通的LB板可以被取代。 <ol><li>称取5毫升去离子水5.5克XLD琼脂和重悬。 </li><li>调匀，直到所有的琼脂是湿的。添加95毫升去离子水，直到所有肿块消失，介质完全悬浮。 </li><li>熬中期至完全溶解（不要高压灭菌）。 </li>...

<ol>
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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=Functional+genomic+analysis+of+C.+elegans+chromosome+I+by+systematic+RNA+interference.">Functional genomic analysis of C. elegans chromosome I by systematic RNA interference.</a> Nature. 408, 325-330 (2000).</li><li>Stiernagle, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maintenance+of+C.+elegans.">Maintenance of C. elegans.</a> WormBook: the online review of C elegans biology. , 1-11 (2006).</li><li>Aballay, A., Ausubel, F. 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线虫是一种简单的遗传模型有机体，吃细菌的营养源。因此，很容易与肠道病原体替代它的正常细菌的食物来调查C之间的相互作用线虫和所选择的病原体。在此协议被描述为沙门氏菌感染和C结合线虫喂食的RNAi治疗研究宿主基因在抵御沙门氏菌感染的作用。曾患协议公开C.线虫蠕虫致病菌他们的一生20时，包括沙门氏菌 。在本?...

自由生活的土壤线虫是用于研究许多生物学问题的简单和基因听话的模式生物。C.线虫显性存在自我施肥雌雄同体。雄性自发地由X染色体的过程中配子1,2非分离产生。在丰富的食物，C的存在线虫通过四个幼虫到成虫不断发展。温度也影响C。线虫的发展;更快的发展在较高温度下观察。在实验室中，C.线虫是培养在20℃下在琼脂平板上的标准温度与晶种菌大肠杆菌 （菌株OP50）作为食物1,2。 在过去的十年中，C.线虫已经成为一个无脊椎生物，研究宿主-病原体相互作用3-5。在自然界中，C.线虫吃细菌的养分吨源1,2。它的正常细菌实验室食品，OP50，可以很容易地被其他病原体检查C之间的相互作用线虫和任何选定的病原体。在这些条件下，肠道是感染的主要部位。事实上，广泛的细菌病原体已经显示出致死性感染C.线虫 3-5。 对革兰阴性菌沙门氏菌是一种胃肠道病原体引起人类食源性疾病的全球6,7。C.线虫是一种很好的模式主机沙门氏菌为这种细菌复制和呈现持续肠道感染8-10。C。线虫已经被用于识别新的和先前已知的沙门氏菌毒力因子11。有趣的是，C。线虫的免疫系统成功地限制了沙门氏菌复制。据此前报道，Inhib所自噬基因银行足球比赛呈现在C增加沙门氏菌复制线虫 ，导致受感染蠕虫10的过早死亡。巨自噬（本文中被称为自噬）是一个涉及亚细胞膜的重排螯合细胞质和细胞器运送到溶酶体降解12动态过程。自噬作用已有报道，以限制沙门氏菌复制在C线虫和哺乳动物10,13。 该三线虫基因组测序的第一个多细胞真核生物的基因组;它是响应于RNAi的治疗14-16。此外，RNA干扰可以被有效地通过使蠕虫摄取含有靶基因，被称为RNA干扰喂食16,17的双链RNA的细菌给药。已经生成的全基因组RNAi筛选16,18全基因组RNAi的喂养库。在本文中， 沙门氏菌感染亲母育酚结合的RNAi喂养，让测试C。感兴趣的线虫基因的防止沙门氏菌感染的能力。

<table border="0" cellpadding="0" cellspacing="0" width="621">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">LB Broth</td>
			<td style="width:119px;">Fisher</td>
			<td style="width:119px;">BP9723-500</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">XLD agar</td>
			<td>EMD Chemicals</td>
			<td>1.05287.0500</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Bacto Agar</td>
			<td>Fisher</td>
			<td>DF0140-01-0</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Peptone</td>
			<td>Fisher</td>
			<td>BP1420-500</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium Chloride</td>
			<td>Fisher</td>
			<td>S671-500</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Calcium Chloride</td>
			<td>Fisher</td>
			<td>C69-500</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Magnesium Sulfate</td>
			<td>Fisher</td>
			<td>M65-500</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">IPTG</td>
			<td>Gold Biotechnology</td>
			<td>12481C50</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cholesterol</td>
			<td>Sigma</td>
			<td>C8667-25G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ampicillin</td>
			<td>Fisher&#160;</td>
			<td>BP1760-25</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Salmonella typhimurium</td>
			<td>ATCC</td>
			<td>ATCC14028</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Petri Dish 95 x 15mm</td>
			<td>Fisher</td>
			<td>FB0875714G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Petri Dish 60 x 15mm&#160;</td>
			<td>Fisher</td>
			<td>08-757-13A&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Falcon Serological pipet</td>
			<td>Fisher</td>
			<td>13-668-2</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Falcon Express Pipet-Aid</td>
			<td>Fisher</td>
			<td>13-675-42</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">MaxQ6000 shaking incubator&#160;</td>
			<td>Thermo Scientific</td>
			<td>SHKE6000-7</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Incubator</td>
			<td>Percival</td>
			<td>I-36DL</td>
			<td></td>
		</tr>
	</tbody>
</table>

a protocol to infect caenorhabditis elegans with salmonella typhimurium

在过去的十年中，C.线虫已经成为一个无脊椎生物，研究宿主和病原体之间的相互作用，包括对革兰阴性菌鼠伤寒沙门氏菌的宿主防御。沙门氏菌建立持续性感染C.肠道线虫和结果在受感染动物的过早死亡。许多免疫机制C.已经确定对线虫对沙门氏菌感染捍卫。吞噬，进化上保守的溶酶体降解途径，已经示出，以限制沙门氏菌复制在C线虫和哺乳动物。这里，一个协议描述来感染C.线虫与鼠伤寒沙门氏菌 ，其中蠕虫暴露在沙门氏菌在有限的时间，类似沙门氏菌感染的人类。 沙门氏菌感染显著缩短C的寿命线虫&lt;/ em&gt;的。使用必需的自噬基因BEC-1作为一个例子，我们结合这种感染的方法与C线虫 RNAi的喂养方法，并表明该协议可以用于检查C的功能线虫的宿主基因在抵御沙门氏菌感染。由于C。线虫全基因组的RNAi文库的情况下，该协议使得它可以全面筛查C。线虫基因，对沙门氏菌等肠道致病菌保护利用全基因组的RNAi文库。

在20°C时，野生型N2蠕虫的寿命中位数为17天（ 图2A和表2）。 沙门氏菌感染显著降低N2蠕虫的寿命中位数天至10.5天（P = 0.0002，log-rank检验）（ 图2A ）。 如果C。线虫基因在抵御沙门氏菌感染中起重要作用，据预测，其抑制将传授易患沙门氏菌感染。事实上，相对于沙门氏菌感染控制的RNAi治疗N2动物的沙...

Watch this Scientific Journal Video about A Protocol to Infect Caenorhabditis elegans with Salmonella typhimurium at JoVE.com

A Protocol to Infect Caenorhabditis elegans with Salmonella typhimurium

一个协议感染<em&gt;秀丽隐杆线虫</em&gt;与<em&gt;鼠伤寒沙门氏菌</em

线虫已成为研究宿主-病原体相互作用的新遗传模型。在这里，我们描述了一个协议，感染C。线虫与鼠伤寒沙门氏菌加上双链RNA干扰干扰技术研究宿主基因在抵御沙门氏菌感染的作用。

In the last decade, C. elegans has emerged as an invertebrate organism to study interactions between hosts and pathogens, including the host defense ...

Research

JoVE Journal

Immunology and Infection

A Protocol to Infect Caenorhabditis elegans with Salmonella typhimurium

An Allelotyping PCR for Identifying Salmonella enterica serovars Enteritidis, Hadar, Heidelberg, and Typhimurium

An Allelotyping PCR for Identifying Salmonella enterica serovars Enteritidis, Hadar, Heidelberg, and Typhimurium

一个用于标识Allelotyping PCR扩增沙门氏菌肠炎血清型，哈达尔，海德堡和鼠伤寒沙门氏菌

Current commercial PCRs tests for identifying Salmonella target genes unique to this genus.  However, there are two species, six subspecies, and over ...

科研

免疫与感染

High-throughput Assay to Phenotype Salmonella enterica Typhimurium Association, Invasion, and Replication in Macrophages

High-throughput Assay to Phenotype Salmonella enterica Typhimurium Association, Invasion, and Replication in Macrophages

高通量检测到的表型<em&gt;沙门氏菌</em&gt;伤寒协会，侵袭和复制的巨噬细胞

Salmonella species are zoonotic pathogens and leading causes of food borne illnesses in humans and livestock1. Understanding the mechanisms underlying ...

Isolation of Salmonella typhimurium-containing Phagosomes from Macrophages

Isolation of Salmonella typhimurium-containing Phagosomes from Macrophages

巨噬细胞中含有吞噬的鼠伤寒沙门氏菌的分离

Salmonella typhimurium is a facultative intracellular bacterium that causes gastroenteritis in humans. After invasion of the lamina propria, S. ...

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

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

在大肠杆菌中生成特异的乙酰蛋白的简易协议

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

A High-throughput Platform for the Screening of Salmonella spp./Shigella spp.

A High-throughput Platform for the Screening of Salmonella spp./Shigella spp.

沙门氏菌筛选的高通量平台.

Fecal-oral transmission of acute gastroenteritis occurs from time to time, especially when people who handled food and water are infected by Salmonella ...

Evaluating Virulence and Pathogenesis of Aeromonas Infection in a Caenorhabditis elegans Model

Evaluating Virulence and Pathogenesis of Aeromonas Infection in a Caenorhabditis elegans Model

线虫模型中气单胞菌感染的毒力及发病机制评价

The human pathogen Aeromonas has been clinically shown to cause gastroenteritis, wound infections, septicemia, and urinary tract infections. Most human ...

Application of Consistent Massage-Like Perturbations on Mouse Calves and Monitoring the Resulting Intramuscular Pressure Changes

在小鼠小腿上应用一致的按摩样扰动，并监测由此产生的肌肉压力变化

Massage is generally recognized to be beneficial for relieving pain and inflammation. Although previous studies have reported anti-inflammatory effects of ...

Using Human Induced Pluripotent Stem Cell-derived Intestinal Organoids to Study and Modify Epithelial Cell Protection Against Salmonella and Other Pathogens

Using Human Induced Pluripotent Stem Cell-derived Intestinal Organoids to Study and Modify Epithelial Cell Protection Against Salmonella and Other Pathogens

利用人类诱导的多能干细胞衍生的肠道有机体研究和改性上皮细胞对沙门氏菌和其他病原体的保护

The intestinal &#8216;organoid&#8217; (iHO) system, wherein 3-D structures representative of the epithelial lining of the human gut can be produced from ...

Chronic Salmonella Infection Induced Intestinal Fibrosis

慢性沙门氏菌感染诱发肠纤维化

Tissue fibrosis characterized by the pathological accumulation of extracellular matrix such as collagen is the outcome of persistent inflammation and ...

NF-&#954;B-dependent Luciferase Activation and Quantification of Gene Expression in Salmonella Infected Tissue Culture Cells

NF-ÃƒÅ½Ã‚ÂºB-dependent Luciferase Activation and Quantification of Gene Expression in Salmonella Infected Tissue Culture Cells

在沙门氏菌感染组织培养细胞中，NF-βB依赖性路西酶的激活和基因表达的定量

The dimeric transcription factor NF-&#954;B regulates many cellular response pathways, including inflammatory pathways by inducing the expression of ...