Name: injection of an. stephensi embryos to generate malaria-resistant mosquitoes
Uploaded: 2007-07-04T00:00:00
Description: Watch this Scientific Journal Video about Injection of An. stephensi Embryos to Generate Malaria-resistant Mosquitoes at JoVE.com

<p class="jove_title">マイクロインジェクションの事前準備： </p><ol><li>血液の供給蚊：月曜日から前の金曜日の水曜日のフィード女性の注射用。木曜日と金曜日を注入するために、同じ週の月曜日にフィード女性。 <br /><br /></li><li>プログラム2と等張緩衝液を用いて石英の針を準備する（資料を参照）。 <br /><br /></li><li>胚は、&quot;管を敷設する&quot;定期的なショウジョウバエ培養バイアルです。それを覆うフィル?...

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
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		<tbody>
		
<tr>
<td>Injection Quartz needles</td>
<td>&nbsp;</td>
<td>Sutter Instrument</td>
<td>&nbsp;</td>
<td>O.D.- 1.0 mm, I.D.-0.70, 10 cm length, are prepared by drawing capillary tubing into a fine 3-8 um with a shaft of approximately 100-300 mm micropipette puller. </td>
</tr>
<tr>
<td>Quartz puller</td>
<td>Tool</td>
<td>Sutter Instrument Co</td>
<td>P-2000</td>
<td>Set-up: Heat-275, Fil-3, Vel-38, Del-250, Pull-141</td>
</tr>
<tr>
<td>EndoFree plasmid kit</td>
<td>&nbsp;</td>
<td>Quiagen</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Isopropanol</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Millex-GV column</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>SLGV RO4 NL</td>
<td>for DNA cleaning</td>
</tr>
<tr>
<td>Construct DNA</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>0.5 mg/ml </td>
</tr>
<tr>
<td>Helper plasmid</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>0.3 mg/ml </td>
</tr>
<tr>
<td>Injection solution</td>
<td>Buffer</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>1x solution: 5mM KCl, 0.1mM sodium phosphate, ph 6. 8</td>
</tr>
<tr>
<td>Isotonic buffer</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>see recipe in the protocol section</td>
</tr>
<tr>
<td>Dessication oil</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Halocarbon oil 700:Halocarbon oil 27(1:1). Mix well. Prepare in advance of microinjection.</td>
</tr>
<tr>
<td>Blood</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>to feed mosquitos</td>
</tr>
<tr>
<td>Anopheles stephensi</td>
<td>Animal</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Mosquitos, male and female.</td>
</tr>
<tr>
<td>Drosophila culture vial</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>for egg laying</td>
</tr>
<tr>
<td>Petri dishes</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>with isotonic buffer, for embryo hatching</td>
</tr>
<tr>
<td>Fine paintbrush</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Sable # 0000</td>
</tr>
<tr>
<td>3MM Whatman paper</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Forceps #5</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>double sided tape</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Microloader</td>
<td>&nbsp;</td>
<td>Eppendorf</td>
<td>5242 956.003</td>
<td>&nbsp;</td>
<tr>
<td>Transjector</td>
<td>&nbsp;</td>
<td>Eppendorf</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Microscope</td>
<td>&nbsp;</td>
<td>Leica</td>
<td>&nbsp;</td>
<td>with moving stage and  micromanipulator, set at 10x magnification </td>
</tr>		</tbody>
		</table>
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injection of an. stephensi embryos to generate malaria-resistant mosquitoes

<p class="jove_content">蚊のゲノムへの外来遺伝子の導入は、関心のある特定の種に合わせたマイクロインジェクションの技術が必要です。このビデオプロトコルが変換された蚊の世代のためのハマダラカの胚にDNA構築物を顕微注入するためにジェームズ実験室で使用されるメソッドを示しています。 、マイクロインジェクションの針を準備する胚を収集し、準備やマイクロインジェクションを行うためのテクニックが示されている。</p>

Watch this Scientific Journal Video about Injection of An. stephensi Embryos to Generate Malaria-resistant Mosquitoes at JoVE.com

Injection of An. stephensi Embryos to Generate Malaria-resistant Mosquitoes

<p class="jove_content">ハマダラカ蚊は、インドやアジア各地に生息するマラリアのためのベクトルです。このビデオでは、蚊にマラリアに抵抗性を付与される導入遺伝子でこの種のmicroinjectionsを実行するためのテクニックを示しています。このビデオで示されて方法論の多くは、他の蚊の種のマイクロインジェクション技術に適用可能である。</p>

の注入。ハマダラカの胚は、マラリア耐性の蚊を生成する

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トランスジェニック蚊を取得するA.ネッタイシマカの胚のマイクロインジェクション

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遺伝子機能を解析するためにゼブラフィッシュ胚のマイクロインジェクション

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Genetic screening is one of the most powerful methods available for gaining insights into complex biological process 1. Over the years many improvements ...

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イメージング<em> C.エレガンス</em落射蛍光顕微鏡とオープンソースソフトウェアを使用して>胚

Cellular processes, such as chromosome assembly, segregation and cytokinesis,are inherently dynamic. Time-lapse imaging of living cells, using ...

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アン<em> in vivoで</emからのタンパク質複合体を分離するために>架橋アプローチ<em>ショウジョウバエ</em>胚

Many cellular processes are controlled by multisubunit protein complexes. Frequently these complexes form transiently and require native environment to ...

A Multi-detection Assay for Malaria Transmitting Mosquitoes

マラリア蚊。送信のためのマルチ検出分析

The Anopheles gambiae species complex includes the major malaria transmitting mosquitoes in Africa. Because these species are of such medical importance, ...

Protocols for Testing the Toxicity of Novel Insecticidal Chemistries to Mosquitoes

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New classes of insecticides with novel modes of action are needed to control insecticide resistant populations of mosquitoes that transmit diseases such ...

Forced Salivation As a Method to Analyze Vector Competence of Mosquitoes

蚊のベクトル能力を分析する方法として唾液の分泌を強制

Vector competence is defined as the potential of a mosquito species to transmit a mosquito-borne virus (mobovirus) to a vertebrate host. Viable virus ...