Name: generation of transgenic c. elegans by biolistic transformation
Uploaded: 2010-08-23T17:00:00
Description: Watch this Scientific Journal Video about Generation of Transgenic C. elegans by Biolistic Transformation at JoVE.com

这项工作是从美国匹兹堡大学和美国国立卫生研究院授予AG028977 ALF的种子资金支持

概述传统的转基因蠕虫所产生的 C.显微注射到转基因DNA 线虫生殖1,2,3,4。虽然成功，这种方法需要专门的设备和经验与蠕虫解剖和显微注射技术的发展。最近枪法已发展作为另一种方法，它使用DNA包覆的金颗粒引入外源DNA的生殖细胞 5,6 。这种方法需要较少的时间在实践方面的投资，成为成功的。 C.枪法线虫生成?...

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基因枪法是一个简单的的方法，引入外源DNA的许多生物，包括 C。 线虫 1,5,6,7,16。依靠黄金颗粒，形成了一个复杂的DNA 与氯化钙存在。阳离子多胺，如亚精胺，保护核酸在体内降解的DNA。既然精胺是一种不稳定的分子，它是重要的存储在小分装于-20 ° C，并在执行轰击之前做出正确的解决方案。正如我们在轰炸协议描述，鱼精蛋白可以用来代替亚精胺提供外源DNA 1...

The volume for a solution in, <a href="http://www.jove.com/video/2090">Generation of Transgenic C. elegans by Biolistic Transformation</a>, was incorrect.

Erratum: Generation of Transgenic C. elegans by Biolistic Transformation

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<td>Gold particles</td>
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<td> P4505</td>
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generation of transgenic c. elegans by biolistic transformation

使用自由生活线虫C.的实验室的数量线虫是迅速增长。这种生物模型的普及是由于快速的一代人的时间和寿命短，维护容易和便宜，完全测序的基因组，和RNAi的资源和突变的动物数组。此外，分析了C.线虫基因组中发现了蠕虫和高等脊椎动物之间极大的相似性，这表明蠕虫病毒的研究可能是一个重要的辅助手段，在整个小鼠培养细胞进行的研究。一个强大的蠕虫病毒的研究和重要组成部分，是利用转基因动物的研究，基因定位和功能的能力。可以创建可以通过蠕虫生殖显微注射，或通过利用基因枪法转基因动物。炮击是一个较新的技术并不太熟悉的实验室。在这里，我们描述了一个简单的协议生成的Bio - Rad公司PDS - 1000系统使用的金粒子枪法转基因蠕虫。到雌雄同体生殖与DNA显微注射法相比，该协议已经不需要特殊技能，从运营商方面识别蠕虫解剖或执行显微注射​​的优势。通常从一个单一的轰炸进一步多个转基因株系。此外，在对比，以显微注射法，基因枪法染色体外的阵列和集成的转基因与转基因动物。能够获得综合的转基因株系，可避免使用整合外源DNA诱变协议。总之，基因枪法可以代转基因动物的一个极具吸引力的一种方法，尤其是对不感兴趣的投资需要的时间和精力，成为显微注射技术熟练的调查。

Watch this Scientific Journal Video about Generation of Transgenic C. elegans by Biolistic Transformation at JoVE.com

Generation of Transgenic C. elegans by Biolistic Transformation

转基因蠕虫常用<em&gt; C。线虫</em&gt;研究。描述的是一个简单而有效，协议引入蠕虫使用DNA包覆的金粒子枪法转基因。参与的努力和轰炸的结果相媲美注射代转基因动物。

生成的基因 C。线虫通过基因枪转化

The number of laboratories using the free living nematode C. elegans is rapidly growing. The popularity of this biological model is attributed to a rapid ...

Research

JoVE Journal

Biology

Generation of Transgenic C. elegans by Biolistic Transformation

Generation of Stable Transgenic C. elegans Using Microinjection

产生稳定的转基因C.使用微量注射线虫

Transgenic Caenorhabditis elegans can be readily created via microinjection of a DNA plasmid solution into the gonad 1.  The plasmid DNA rearranges to ...

科研

生物学

Paradigms for Pharmacological Characterization of C. elegans Synaptic Transmission Mutants

为线虫突触传递的突变体的药理特性的范式

The nematode, Caenorhabditis elegans, has become an expedient model for studying neurotransmission. C. elegans is unique among animal models, as the ...

Dissecting and Recording from The C. Elegans Neuromuscular Junction

线虫神经肌肉交界处的解剖和录制

Neurotransmission is the process by which neurons transfer information via chemical signals to their post-synaptic targets, on a rapid time scale.   This ...

Generation of RNA/DNA Hybrids in Genomic DNA by Transformation using RNA-containing Oligonucleotides

改造使用含RNA寡核苷酸基因组DNA的RNA / DNA杂交一代

Synthetic short nucleic acid polymers, oligonucleotides (oligos), are the most functional and widespread tools of molecular biology. Oligos can be ...

Basic Caenorhabditis elegans Methods: Synchronization and Observation

Basic Caenorhabditis elegans Methods: Synchronization and Observation

基本的<em&gt;线虫</em&gt;方法：同步和观察

Research into the molecular and developmental biology of the nematode Caenorhabditis elegans was begun in the early seventies by Sydney Brenner and it has ...

C. elegans Chemotaxis Assay

C. elegans Chemotaxis Assay

线虫趋含量的

Many organisms use chemotaxis to seek out food sources, avoid noxious substances, and find mates.  Caenorhabditis elegans has impressive chemotaxis ...

A Method for Culturing Embryonic C. elegans Cells

A Method for Culturing Embryonic C. elegans Cells

一种用于培养胚胎<em&gt; C。线虫</em&gt;细胞

C. elegans is a powerful model system, in which genetic and molecular techniques are easily applicable. Until recently though, techniques that require ...

Antibody Staining in C. Elegans Using &quot;Freeze-Cracking&quot;

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抗体在染色<em&gt; C。线虫</em&gt;使用“冻结开裂”

To stain C.  elegans with antibodies, the relatively impermeable cuticle must be  bypassed by chemical or mechanical methods.  "Freeze-cracking" is one ...

An Anoxia-starvation Model for Ischemia/Reperfusion in C. elegans

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对缺血/再灌注在缺氧，饥饿型号<em&gt; C。线虫</em

Protocols for anoxia/starvation in the genetic model organism C. elegans simulate ischemia/reperfusion. Worms are separated from bacterial food and placed ...

Culturing Caenorhabditis elegans in Axenic Liquid Media and Creation of Transgenic Worms by Microparticle Bombardment

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培养<em&gt;秀丽隐杆线虫</em &gt;在无菌液体介质与创造转基因蠕虫通过微粒轰击

In this protocol, we present the required materials, and the procedure for making modified&#160;C. elegans&#160;Habituation and Reproduction media ...