Name: complementary use of microscopic techniques and fluorescence reading in studying cryptococcus-amoeba interactions
Uploaded: 2019-06-22T13:00:00
Description: Watch this Scientific Journal Video about Complementary Use of Microscopic Techniques and Fluorescence Reading in Studying Cryptococcus-Amoeba Interactions at JoVE.com

这项工作得到了南非国家研究基金会(赠款号:87903年)和自由州大学的赠款的支持。我们也感谢彼得·范·怀克和汉莉·格罗布勒在我们的显微镜研究中提供的服务和协助。

隐球新毒杆菌和一些阿坎塔莫埃巴卡斯特兰尼菌株被视为生物安全-2级(BSL-2)病原体;因此,研究人员在使用这些生物时必须采取适当的预防措施。例如,实验室人员应接受专门的培训和个人防护设备 (PPE),如实验室外套、手套和眼睛防护。生物安全柜(2级)应用于可能导致感染的程序14。
1. 真菌细胞的培养和标准化(从Madu等人改性15)
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在本文中,成功地使用了不同的技术来揭示当阿米巴与隐球菌细胞相互作用时可能出现的结果。此外,我们有兴趣展示3-羟基脂肪酸对隐球菌-阿米巴相互作用结果的影响。
第一种技术是共聚焦显微镜,它渲染静止图像。此处此技术的主要缺点是,它只为我们提供了仅限于特定时间点的信息。任何可以根据结果得出的结论都有助于归纳推理,其中人们可以根据一组观察<sup class="...

微生物经过一段时间的进化,在不同的生态区中占据并茁壮成长,如土壤和水的开放物理边界,等等。在这些利基中,微生物往往直接争夺有限的资源;重要的是,对于营养,他们用来支持他们的生长或空间,他们需要容纳不断扩大的人口2,3。在某些情况下,一些霍洛索生物,如阿米巴甚至可能早于隐球细胞,作为从生物量4,5中提取营养物质的一种方式。反过来,这允许这些生物通过控制猎物的数量来确立领土优势。由于这种掠夺性压力,一些猎物可能被选择来产生微生物因子,如隐球胶囊6,以调和压力的负面影响。然而,作为这种压力的意外后果,一些微生物获得一些因素,使它们能够跨越物种屏障,寻找新的利基来殖民7,如人体的封闭空间,富含营养,具有理想条件。后者可以解释陆地微生物如何像隐球菌(C.新福曼人可以转化为致病性。
为此,必须研究隐球细胞与阿米巴的初始接触,以及如何选择它们成为致病菌。更具体地说,这可能为隐球细胞在感染期间受到巨噬细胞作用时的行为提供线索。正是出于这个原因,阿米巴被选为这里巨噬细胞的模型,因为它是相对便宜和容易保持阿米巴文化在实验室8。感兴趣的也是研究隐球菌二次代谢物(即3-羟基脂肪酸9,10)如何影响阿米巴和隐球细胞之间的相互作用。
用肉眼感知阿米巴和猎物之间的相互作用的一种简单方法是在琼脂盘表面使用猎物创建草坪,并发现阿米巴。琼脂板上的斑块或透明区域的可视化描绘了阿米巴可能以猎物为食的区域。然而,在这个宏观层面上,只注意到这一进程的结果,并且不能观察到噬菌体过程是机械化的。因此,为了在细胞到细胞的基础上欣赏这个过程,有几种微观方法可以使用11,12。例如,带孵化室的倒置显微镜可用于视频记录噬菌细胞与其目标13之间的事件延时。不幸的是,由于具有延时功能的显微镜的成本,实验室并不总是能够购买这种显微镜,尤其是在资源贫乏的环境中。
为了规避上述限制,本研究提出了一个顺序探索设计,评估C.新福曼斯(C.新福曼斯UOFS Y-1378)和C.新福曼斯LMPE 046与阿坎塔莫埃巴·卡斯特拉尼的相互作用.首先,使用在定量方法之前的定性方法。静止图像使用倒置荧光显微镜以及透射电子显微镜拍摄,以描绘阿米巴-隐球菌相互作用。随后,使用板读取器对荧光进行量化,以估计阿米巴将隐球细胞内化的效率。在数据解释阶段协调这些方法的发现时,这可能与浏览方位体延时视频一样多的关键信息。

<table border="0" cellpadding="0" cellspacing="0" style="width:783px;" width="782">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">1,4-Diazabicyclo-[2.2.2]-octane</td>
			<td style="width:176px;">Sigma-Aldrich</td>
			<td style="width:123px;">D27802</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">1.5-mL plastic tube&#160;</td>
			<td>Thermo Fisher Scientific</td>
			<td style="width:123px;">69715</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">15-mL Centrifuge tube&#160;</td>
			<td>Thermo Fisher Scientific</td>
			<td style="width:123px;">7252018</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">50-mL Centrifuge tube&#160;</td>
			<td>Thermo Fisher Scientific</td>
			<td style="width:123px;">1132017</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">8-Well chamber slide</td>
			<td style="width:176px;">Thermo Fisher Scientific</td>
			<td style="width:123px;">1109650</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">Acetone</td>
			<td style="width:176px;">Merck</td>
			<td style="width:123px;">SAAR1022040LC</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:312px;">Amoeba strain</td>
			<td style="width:176px;">ATCC&#210;</td>
			<td style="width:123px;">30234TM</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:312px;">ATCC medium 712</td>
			<td style="width:176px;">ATCC&#210;</td>
			<td style="width:123px;">712TM</td>
			<td style="width:172px;">Amoeba medium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Black 96-well microtiter plate</td>
			<td>Thermo Fisher Scientific</td>
			<td style="width:123px;">152089</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">Centrifuge</td>
			<td style="width:176px;">Hermle</td>
			<td style="width:123px;">-</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">Chloroform</td>
			<td style="width:176px;">Sigma-Aldrich</td>
			<td style="width:123px;">C2432</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">Confocal microscope</td>
			<td style="width:176px;">Nikon</td>
			<td>Nikon TE 2000</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td colspan="4" height="21" style="height:21px;width:783px;">Epoxy resin:</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">[1] NSA</td>
			<td style="width:176px;">[1] ALS</td>
			<td style="width:123px;">[1] R1054</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">[2] DER 736</td>
			<td style="width:176px;">[2] ALS</td>
			<td style="width:123px;">[2] R1073</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">[3] ERL Y221 resin</td>
			<td style="width:176px;">[3] ALS</td>
			<td style="width:123px;">[3] R1047R</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">[4] S1 (2-dimethylaminoethanol)</td>
			<td style="width:176px;">[4] ALS&#160;</td>
			<td style="width:123px;">[4] R1067</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">Fluorescein isothiocyanate</td>
			<td style="width:176px;">Sigma-Aldrich</td>
			<td style="width:123px;">F4274</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">Formic Acid</td>
			<td style="width:176px;">Sigma-Aldrich</td>
			<td style="width:123px;">489441</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fluoroskan Ascent FL</td>
			<td>Thermo Fisher Scientific</td>
			<td style="width:123px;">374-91038C</td>
			<td style="width:172px;">Microplate reader</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">Glucose</td>
			<td style="width:176px;">Sigma-Aldrich</td>
			<td style="width:123px;">G8270</td>
			<td>-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">Glutaraldehyde</td>
			<td style="width:176px;">ALS</td>
			<td style="width:123px;">R1009</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">Hemocytometer</td>
			<td style="width:176px;">Boeco</td>
			<td style="width:123px;">-</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">Lead citrate</td>
			<td style="width:176px;">ALS</td>
			<td style="width:123px;">R1209</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Liquid Chromatography Mass Spectrometer</td>
			<td>Thermo Fisher Scientific</td>
			<td style="width:123px;"></td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">Methanol</td>
			<td style="width:176px;">Sigma-Aldrich</td>
			<td style="width:123px;">R 34,860</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">Orbital shaker</td>
			<td style="width:176px;">Lasec&#160;</td>
			<td style="width:123px;">-</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">Osmium tetroxide</td>
			<td style="width:176px;">ALS</td>
			<td style="width:123px;">R1015</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:312px;">pHrodo Green Zymosan A BioParticles</td>
			<td style="width:176px;">Life Technologies</td>
			<td style="width:123px;">P35365</td>
			<td style="width:172px;">This is the pH-sensitive dye</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">Physiological buffer solution</td>
			<td style="width:176px;">Sigma-Aldrich</td>
			<td style="width:123px;">P4417-50TAB</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">Rotary shaker</td>
			<td style="width:176px;">Labcon</td>
			<td style="width:123px;">-</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td colspan="4" height="21" style="height:21px;width:783px;">Sodium phosphate buffer:</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:312px;">[1] di-sodium hydrogen orthophosphate dihydrate&#160;</td>
			<td style="width:176px;">[1] Merck</td>
			<td style="width:123px;">[1] 106580</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:312px;">[2] sodium di-hydrogen orthophosphate dihydrate</td>
			<td style="width:176px;">[2] Merck&#160;</td>
			<td style="width:123px;">[2] 106345</td>
			<td style="width:172px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">Transmission electron microscope</td>
			<td style="width:176px;">Philips</td>
			<td>Philips EM 100&#160;</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">Trypan blue&#160;</td>
			<td style="width:176px;">Sigma-Aldrich</td>
			<td style="width:123px;">T8154</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">Ultramicrotome</td>
			<td style="width:176px;">Leica</td>
			<td style="width:123px;">EM UC7</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">Uranyl acetate</td>
			<td style="width:176px;">ALS</td>
			<td style="width:123px;">R1260A</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">Vacuum dessicator</td>
			<td style="width:176px;">Lasec&#160;</td>
			<td style="width:123px;">-</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">Vial</td>
			<td style="width:176px;">Sigma-Aldrich</td>
			<td style="width:123px;">29651-U</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">YNB</td>
			<td style="width:176px;">Lasec&#160;</td>
			<td style="width:123px;">239210</td>
			<td style="width:172px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:312px;">YPD agar</td>
			<td style="width:176px;">Sigma-Aldrich</td>
			<td style="width:123px;">Y-1500</td>
			<td>-</td>
		</tr>
	</tbody>
</table>

complementary use of microscopic techniques and fluorescence reading in studying cryptococcus-amoeba interactions

为了模拟隐球菌感染,阿米巴是环境中隐球细胞的天然捕食者,可以作为巨噬细胞的模型。这种食肉生物,类似于巨噬细胞,利用噬菌体杀死内化细胞。借助共聚焦激光扫描显微镜,可以捕获描述隐球细胞和阿米巴之间交互时刻的图像。电子显微镜的分辨率也有助于揭示隐球细胞的超结构细节,当被困在阿米巴食物真空中时。由于噬菌体是一个连续的过程,因此定量数据随后被集成到分析中,以解释捕获图像的时间点时发生的情况。具体来说,读取相对荧光单位是为了量化阿米巴在隐球细胞内化中的效率。为此,隐球细胞被染色的染料,使他们荧光一旦被困在食物的酸性环境中。当一起使用时,通过此类技术收集的信息可以提供关键信息,以帮助得出细胞在阿米巴内部化时以及可能由其他噬菌体细胞内化时的行为和命运的结论。

微生物是肉眼无法感知的微生物。然而,它们的影响可能导致可观察到的临床明显疾病,如皮肤感染。在研究微生物的某些方面时,从微生物的形态、副产品和相互作用,能够提供图片和视频证据至关重要。
我们首先试图可视化隐球细胞和阿米巴之间的相互作用。为此,首先研究了显示2小时共育细胞的明亮场图像。一张图片显示一?...

Watch this Scientific Journal Video about Complementary Use of Microscopic Techniques and Fluorescence Reading in Studying Cryptococcus-Amoeba Interactions at JoVE.com

Complementary Use of Microscopic Techniques and Fluorescence Reading in Studying Cryptococcus-Amoeba Interactions

在研究隐球菌-阿米巴相互作用中,显微技术和荧光读数的补充应用

本文详细介绍了利用静止光图像和高分辨率透射电子显微镜图像进行制备隐球细胞和阿米巴共同培养的协议。此处说明定量数据如何补充此类定性信息。

To simulate Cryptococcus infection, amoeba, which is the natural predator of cryptococcal cells in the environment, can be used as a model for ...

该程序的总体目的是利用显微镜作为工具，研究其环境捕食者阿米巴的致病真菌物种隐形球菌新形态的噬菌体。本视频详细介绍了制备隐球细胞和阿米巴联合培养的协议，我将使用共生激光扫描显微镜和传输电子显微镜来研究该协议。从使用这些技术获得的信息可以深入了解在感染期间内化时隐球细胞的行为。在YPD芳球盘上划掉测试真菌菌株。在30摄氏度下孵育板48小时。从48小时厚的盘子里刮掉一个循环的细胞，并接种到含有100毫升YNB汤的250毫升圆锥形烧瓶中，并辅以4%的葡萄糖。在30摄氏度的旋转摇床上孵育烧瓶24小时。经过24小时的孵育期，使用血细胞计计算真菌细胞，并使用生理缓冲液将细胞数调整为每毫升100万个细胞。培养peton酵母中的阿米巴细胞在30摄氏度下提取葡萄糖汤一周。使用血细胞计计算阿米巴细胞，并使用新鲜无菌 ATCC 介质 712 将细胞数调整为每毫升 1000 万细胞。之后，使用锥蓝色污渍进行可行性测定。建议使用至少显示 80% 生存能力的细胞。将标准化阿米巴细胞的 200 微升悬浮液分配到粘附幻灯片的腔室中。让细胞在 30 摄氏度下粘附在表面，需要两个小时。当阿米巴细胞沉淀下来时，用荧光素同位素染色的标准化隐球细胞染色。在室温下和黑暗中轻轻搅动真菌细胞两小时。将染色的隐球细胞洗涤后，将200微升悬浮液分配到已经含有阿米巴细胞的适当腔室。在30摄氏度下培育准备好的共培养，再多培养两个小时。在开投期结束时，用PBS清洗室两次，以去除任何未绑定的阿米巴细胞，包括非内化的隐球细胞。吸气用于修复细胞的谷胱甘肽，用PBS清洗室两次。拆卸腔室滑轨。使用共合激光扫描显微镜查看共培养细胞。为了补充上述测定，用pHrodo染料对隐球细胞进行染色，选择性地允许读取被困在食物气穴内的细胞。将这些染色细胞放入已经含有种子阿米巴细胞的黑色粘附微刺激板的孔中。测量微孔板读取器上的荧光，该显微板读取器可以将对数信号转换为相对荧光单元。将五毫升标准化的隐球细胞添加到已经包含五毫升标准化阿米巴细胞的离心管中。让管子在30摄氏度下站立两小时。离心后吸上经。修复代表共培养细胞的颗粒，用磷酸钠一摩尔在pH 7缓冲3%谷胱甘肽3小时。用磷酸钠缓冲液清洗共培养细胞一次。再次修复1个半小时，在类似的缓冲1%的三氧化硅，然后洗涤。在分级丙酮系列中，TEM材料也被称为共培养细胞，每个 30%50%95%和 100%的 15 分钟。在这里，在两个变化中重复最后的脱水步骤。通过权衡正常稠度环氧树脂来准备环氧树脂。将材料嵌入环氧树脂中。在 70 摄氏度下聚合 TEM 材料 8 小时。用安装在超原子上的玻璃刀切割 16 纳米部分。在黑暗中用醋酸烷染色10分钟，然后用铅柠檬酸盐染色10分钟，也在黑暗中。使用透射电子显微镜查看各部分。为了说明目的，在视频中，检查了两种分离物，一种是产生3-羟基脂肪酸，另一种是不会产生的。 3-羟基脂肪酸是二次代谢物，在隐球细胞的生长中不能发挥作用。下面是显示隐球和阿米巴之间的互动时刻的快照。到点，一个加密球菌细胞之前和之后内化可以观察到。在研究噬菌体时，利用活成像是理想的选择。然而，研究人员并不总是能够获得这种显微镜，以便持续跟踪噬菌体的过程。补偿此限制的可能解决方案可能是通过读取相对荧光单位来补充图像的定量数据。条形图是用于补充图像的数据示例。重要的是，图像形式的定性数据和定量数据都可以在不同时间点拍摄。通过拼凑所有信息，我们可以开始构建一个更大的图像，在吞噬过程中发生的事情。从定量数据中可以清楚地看出，与不产生3-羟基脂肪酸的细胞相比，含有3-羟基脂肪酸的细胞对阿米巴作用的影响较小，因为内化细胞较少。TEM 是一个特别强大的工具，因为它具有高分辨率，它可以让鸟瞰食物气液的流明，这是检查实时成像和静止图像时经常错过的细节。在这个特定的TEM显微图中，可以清楚地看到隐球细胞表面的 protube。此前，理论认为这些细胞表面结构是3-羟基脂肪酸释放到周围环境的通道。到点，这些渠道可以帮助提供3-羟基脂肪酸到阿米巴的食物气液的流明，以改变内部条件，从而促进细胞生存。pHrodo 或像 Fitc 可能是一个更好的污渍使用， 因为它只荧光在低 pH， 像在食物气管内。具体说来，pHrodo只允许研究人员获得有关内化细胞的数据。为此，重要的是细胞在染色前悬浮在磷酸盐缓冲液中，而阿米巴介质是中性pH值的。传输电子显微镜操作员应经过良好的训练，以正确分段，因为这通常是结果可能失真和显微图被电子轰击损坏的点。据设想，研究人员将能够从提出的协议中获得足够的信息，并在他们自己的研究中对其进行优化。此外，研究人员还可以开发针对靶向代谢物的抗体，并在免疫荧光显微镜研究中应用这些抗体，包括在TEM检查期间使用免疫金标签。

Research

JoVE Journal

Immunology and Infection

Complementary Use of Microscopic Techniques and Fluorescence Reading in Studying Cryptococcus-Amoeba Interactions

Use of an Optical Trap for Study of Host-Pathogen Interactions for Dynamic Live Cell Imaging

动态活细胞成像的宿主 - 病原体相互作用的研究使用的光学陷阱

Dynamic live cell imaging allows direct visualization of real-time interactions between cells of the immune system1, 2; however, the lack of spatial and ...

科研

免疫与感染

Use of the EpiAirway Model for Characterizing Long-term Host-pathogen Interactions

使用的EpiAirway模型，用于描述长期宿主 - 病原体相互作用

Nontypeable Haemophilus influenzae  (NTHi) are human-adapted Gram-negative bacteria that can cause recurrent and chronic infections of the respiratory ...

Assessing Hepatic Metabolic Changes During Progressive Colonization of Germ-free Mouse by 1H NMR Spectroscopy

Assessing Hepatic Metabolic Changes During Progressive Colonization of Germ-free Mouse by 1H NMR Spectroscopy

在无菌无鼠标进定植评估肝代谢的变化 1 H核磁共振光谱

It is well known that gut bacteria contribute significantly to the host homeostasis, providing a range of benefits such as immune protection and vitamin ...

Studying Interactions of Staphylococcus aureus with Neutrophils by Flow Cytometry and Time Lapse Microscopy

Studying Interactions of Staphylococcus aureus with Neutrophils by Flow Cytometry and Time Lapse Microscopy

相互作用的研究<em&gt;金黄色葡萄球菌</em与中性粒细胞通过流式细胞仪和时间推移显微镜

We present methods to study the effect of phenol soluble modulins (PSMs)  and other toxins produced and secreted by Staphylococcus  aureus on neutrophils. ...

Use of Shigella flexneri to Study Autophagy-Cytoskeleton Interactions

Use of Shigella flexneri to Study Autophagy-Cytoskeleton Interactions

使用<em&gt;福氏痢疾杆菌</em&gt;研究自噬，细胞骨架相互作用

Shigella flexneri is an intracellular pathogen that can escape from phagosomes to reach the cytosol, and polymerize the host actin cytoskeleton to promote ...

A Novel In vitro Model for Studying the Interactions Between Human Whole Blood and Endothelium

A Novel In vitro Model for Studying the Interactions Between Human Whole Blood and Endothelium

一种新型<em&gt;在体外</em&gt;模型研究人类全血细胞间的相互作用

The majority of all known diseases are accompanied by disorders of the cardiovascular system. Studies into the complexity of the interacting pathways ...

Evaluation of the Efficacy And Toxicity of RNAs Targeting HIV-1 Production for Use in Gene or Drug Therapy

RNA的疗效和毒性的评价基因或药物治疗靶向HIV-1的生产使用

Small RNA therapies targeting post-integration steps in the HIV-1 replication cycle are among the top candidates for gene therapy and have the potential ...

Visual and Microscopic Evaluation of Streptomyces Developmental Mutants

Visual and Microscopic Evaluation of Streptomyces Developmental Mutants

链霉菌发育突变体的视觉和显微评价

Streptomycetes are filamentous soil bacteria belonging to the phylum Actinobacteria that are found throughout the world and produce a wide array of ...

Co-immunoprecipitation Assay for Studying Functional Interactions Between Receptors and Enzymes

免疫沉淀法研究受体与酶的功能相互作用

Receptor-associated enzymes are the major mediators of cellular activation. These enzymes are regulated, at least in part, by physical interactions with ...

Analysis of SAMHD1 Restriction by Flow Cytometry in Human Myeloid U937 Cells

流式细胞术分析人髓系U937细胞中SAMHD1限制性

Sterile &#945;-motif/histidine-aspartate domain-containing protein 1 (SAMHD1) inhibits replication of HIV-1 in quiescent myeloid cells. U937 cells are ...