Name: colonization of euprymna scolopes squid by vibrio fischeri
Uploaded: 2012-03-01T13:00:00
Description: Watch this Scientific Journal Video about Colonization of Euprymna scolopes Squid by Vibrio fischeri at JoVE.com

作者感谢马蒂亚斯鱿鱼设施的支持和迈克尔·哈德菲尔德和Kewalo海洋实验室在现场收集的援助，并为这个协议作出贡献的Ruby和麦克福尔-艺实验室的成员在这个手稿的意见，Gyllborg。在曼德尔实验室的工作是支持由NSF IOS-0843633。

1。细菌菌剂的制备<ol><li> 0天 前两天鱿鱼接种，板13琼脂上的LBS相关的细菌株。 </li><li>在25-28℃过夜孵育细菌。 </li><li> 第1天 接种一五，各殖民地3毫升磅玻璃文化管中等鲵株感染。准备作为备份的重复管。 </li><li> 第2天 （协调细菌步骤与鱿鱼步骤3.7-3.10 1.4-1.6） 1小时前接种，继代培养细菌1:80（37.5μL）...

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	<li>Aas, J. A., Paster, B. J., Stokes, L. N., Olsen, I., Dewhirst, F. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Defining+the+normal+bacterial+flora+of+the+oral+cavity.">Defining the normal bacterial flora of the oral cavity.</a> J. Clin. Microbiol. 43, 5721-5732 (2005).</li><li>Mandel, M. J., Wollenberg, M. S., Stabb, E. V., Visick, K. L., Ruby, E. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+single+regulatory+gene+is+sufficient+to+alter+bacterial+host+range.">A single regulatory gene is sufficient to alter bacterial host range.</a> Nature. 458, 215-218 (2009).</li><li>Grice, E. A., Segre, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+skin+microbiome.">The skin microbiome.</a> Nat. Rev. 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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+winnowing:+establishing+the+squid-Vibrio+symbiosis.">The winnowing: establishing the squid-Vibrio symbiosis.</a> Nat. Rev. Microbiol. 2, 632-642 (2004).</li><li>Ruby, E. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lessons+from+a+cooperative,+bacterial-animal+association:+the+Vibrio+fischeri-Euprymna+scolopes+light+organ+symbiosis.">Lessons from a cooperative, bacterial-animal association: the Vibrio fischeri-Euprymna scolopes light organ symbiosis.</a> Annu. Rev. Microbiol. 50, 591-624 (1996).</li><li>McFall-Ngai, M. J., Ruby, E. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Symbiont+recognition+and+subsequent+morphogenesis+as+early+events+in+an+animal-bacterial+mutualism.">Symbiont recognition and subsequent morphogenesis as early events in an animal-bacterial mutualism.</a> Science. 254, 1491-1494 (1991).</li><li>Jones, B., Nishiguchi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Counterillumination+in+the+Hawaiian+bobtail+squid,+Euprymna+scolopes+Berry+(Mollusca:+Cephalopoda).">Counterillumination in the Hawaiian bobtail squid, Euprymna scolopes Berry (Mollusca: Cephalopoda).</a> Marine Biology. 144, 1151-1155 (2004).</li><li>Graf, J., Ruby, E. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Host-derived+amino+acids+support+the+proliferation+of+symbiotic+bacteria.">Host-derived amino acids support the proliferation of symbiotic bacteria.</a> Proc. Natl. Acad. Sci. U.S.A. 95, 1818-1822 (1998).</li><li>Ruby, E. G., McFall-Ngai, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+squid+that+glows+in+the+night:+development+of+an+animal-bacterial+mutualism.">A squid that glows in the night: development of an animal-bacterial mutualism.</a> J. Bacteriol. 174, 4865-4870 (1992).</li><li>Lee, P. N., McFall-Ngai, M. J., Callaerts, P., de Couet, H. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Hawaiian+bobtail+squid+(Euprymna+scolopes):+a+model+to+study+the+molecular+basis+of+eukaryote-prokaryote+mutualism+and+the+development+and+evolution+of+morphological+novelties+in+cephalopods.">The Hawaiian bobtail squid (Euprymna scolopes): a model to study the molecular basis of eukaryote-prokaryote mutualism and the development and evolution of morphological novelties in cephalopods.</a> Cold Spring Harbor Protocols. , (2009).</li><li>Stabb, E., Visick, K., Millikan, D., Corcoran, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Vibrio+fischeri-Euprymna+scolopes+symbiosis:+a+model+marine+animal-bacteria+interaction.">The Vibrio fischeri-Euprymna scolopes symbiosis: a model marine animal-bacteria interaction.</a> Recent Advances in Marine Science and Technology. , (2001).</li><li>Boettcher, K. 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描述的殖民法，允许在受控的实验室环境中的一种自然的共生过程的分析。因此，它可以用来评估由不同的自然株，突变株，并根据不同的化学制度的殖民化。通常用来描述实验上的变化，以评估不同方面的共生。可以测量殖民动力学检查在第一个24小时，它可以自动检测巧合探测器已被删除，在闪烁计数器的发光。此外，一个应变，相对于其他相对定植能力，可以通过有竞争力的殖民实验测量?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td>玻璃培养管，直径16毫米</td><td>厂商VWR </td><td> 47729-580 </td><td></td></tr><tr><td>上限为玻璃培养管</td><td>费舍尔</td><td> NC9807998 </td><td></td></tr><tr><td>可见分光光度计测定OD值600 </td><td> biowave </td><td> CO8000 </td><td>任何能够测量外径600分光光度计的工作。本机可直接测量液体的OD 600在玻璃培养管。一些接种计算的调整可能是必要的，取决于所使用的仪器。 </td></tr><tr><td> GloMax 20/20单管光度计</td><td> Promega公司</td><td> E5311 </td><td>相当于Turner公司20/20n光度计。包括离心管架。 </td></tr><tr><td> GloMax 20/20晚间标准</td><td> Promega公司</td><td> E5341 </td><td>用于光度计校正。 </td></tr><tr><td>折射仪，手持式</td><td>福斯特和史密斯游泳</td><td> CD-14035 </td><td>校准之前，每次使用去离子水。使用后每用去离子水冲洗，以防止盐积聚。 </td></tr><tr><td>即时海洋（人工海水浓缩液） </td><td>福斯特和史密斯游泳</td><td> CD-16881 </td><td>准备在35‰，在去离子水，使用的验光仪，通过SFCA 0.2微米的过滤器，然后过滤。 </td></tr><tr><td>过滤装置</td><td> NALGENE </td><td> 158-0020 </td><td>表面活性剂醋酸纤维素（SFCA）膜，0.2微米。我们已经观察到一些含表面活性剂的聚醚砜过滤器变量的结果。 </td></tr><tr><td>转移移液器</td><td>费舍尔</td><td> 13-711 -上午9时</td><td>用剪刀或刀片，切干净以上第一山脊增加枪头直径和避免挤压鱿鱼幼体的一角。 </td></tr><tr><td>一次性样品碗（塑料酒杯） </td><td>彗星</td><td> T9S（9盎司）。 </td><td>接种碗，上直径3¼“，直径2¼”，高度为3“。碗创建一个同质的环境，因为他们没有底部的轮辋，其中鱿鱼可以得到被困在低氧气小生。大小进行了优化40毫升接种。可用在webstaurantstore.com，619PI9。 </td></tr><tr><td> 果蝇样品瓶</td><td>厂商VWR </td><td> 89092-720 </td><td>瓶直径相匹配的光电倍增管上的光度计开幕。 </td></tr><tr><td> 1.5毫升离心管</td><td> ISC的Bioexpress </td><td>的C-3217-1CS </td><td>管必须符合杵的形状。 </td></tr><tr><td>乙醇，200证明</td><td>费舍尔</td><td>端口BP2818-100 </td><td></td></tr><tr><td>杵</td><td>金布尔大通/ Kontes </td><td> 749521-1500 </td><td></td></tr><tr><td>电镀珠，直径为5毫米</td><td>金布尔大通</td><td> 13500 5 </td><td>每管和高压灭菌器，准备5。 </td></tr></tbody></table>

colonization of euprymna scolopes squid by vibrio fischeri

与特定的细菌被发现在动物组织1-5。这种细菌主机协会（共生）可能是有害的（致病性），有没有健身的后果（共生），或有利于（互惠）。而备受人们关注已给病原相互作用，知之甚少的进程决定的有利/共生细菌的环境重现收购。轻器官之间的海洋革兰氏阴性菌共生五鲵和夏威夷短尾鱿鱼，E. scolopes，代表着一个非常具体的互动，其中一台主机（E. scolopes）建立贯穿其一生当然6,7只有一个菌种（ 五鲵 ）的共生关系。由五，生产发光鲵在这种互动提供了一个五反掠夺利益在8,9夜间活动scolopes，而营养丰富的宿主组织提供V。鲵与受保护的利基10。在每个主机上一代，这一关系的概括，从而较可预测的过程，可以在共生发展的各个阶段的详细评估。少年鱿鱼孵化aposymbiotically（uncolonized），并在实验室中，如果前30-60分钟内收集和转移到共生的自由水，不能被殖民除实验接种6。因此，这种互动提供了一个有用的模型系统，在其中进行评估，导致从11,12环境的共生微生物的具体收购的各个步骤。 在这里，我们描述了一种方法来评估发生刚出时aposymbiotic 大肠杆菌殖民化的程度scolopes暴露（人造）海水中含有五。鲵。这个简单的实验说明接种，自然感染和恢复E.新生的光器官的细菌共生scolopes。护理提供一个一致的环境共生的发展过程中的动物，尤其是关于水质和轻线索。特征描述的共生人口的方法包括：（1）的细菌派生发光的测量，（2）直接菌落计数恢复共生。

Watch this Scientific Journal Video about Colonization of Euprymna scolopes Squid by Vibrio fischeri at JoVE.com

Colonization of Euprymna scolopes Squid by Vibrio fischeri

方法概括的过程，其中夏威夷短尾鱿鱼，<em&gt; Euprymna scolopes</em&gt;和其细菌的共生体，<em&gt;弧菌鲵</em&gt;，分别提出，然后介绍了允许由细菌鱿鱼轻机关的具体殖民。定植细菌衍生的发光性能和直接菌落计数检测。

定植 Euprymna scolopes鱿鱼由弧菌鲵</em

Specific bacteria are found in association with animal tissue1-5. Such host-bacterial associations (symbioses) can be detrimental (pathogenic), have no ...

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