Name: examination of the telomere g-overhang structure in trypanosoma brucei
Uploaded: 2011-01-26T17:00:00
Description: Watch this Scientific Journal Video about Examination of the Telomere G-overhang Structure in Trypanosoma brucei at JoVE.com

我们想感谢卡罗琳价格博士科学讨论和有见地的建议。这项工作是由美国国立卫生研究院授予AI066095（PI李：碧波）的支持。

1。检测T。布氏端粒G -悬使用母语-凝胶杂交3 原理（图1）：在自然条件下，结束标记（CCCTAA）4（TELC）寡核苷酸探针只能杂交的单链端粒G -悬地区。杂交强度悬的长度成正比。变性，中和后，相同的探测器将能够杂交整个端粒区域。杂交强度代表总端粒DNA量和可用于装载控制。这个实验的两个标准控制杂交使用末端标记（TTAGGG）4（TELG）寡核苷酸...

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	<li>Munoz-Jordan, J. L., Cross, G. A., de Lange, T., Griffith, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=t-loops+at+trypanosome+telomeres.">t-loops at trypanosome telomeres.</a> EMBO J. 20, 579-588 (2001).</li><li>Li, B., Espinal, A., Cross, G. A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trypanosome+telomeres+are+protected+by+a+homologue+of+mammalian+TRF2.">Trypanosome telomeres are protected by a homologue of mammalian TRF2.</a> Mol Cell Biol. 25, 5011-5021 (2005).</li><li>Wellinger, R. J., Wolf, A. J., Zakian, V. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Saccharomyces+telomeres+acquire+single-strand+TG1-3+tails+late+in+S+phase.">Saccharomyces telomeres acquire single-strand TG1-3 tails late in S phase.</a> Cell. 72, 51-60 (1993).</li><li>McElligott, R., Wellinger, R. 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+terminal+DNA+structure+of+mammalian+chromosomes.">The terminal DNA structure of mammalian chromosomes.</a> EMBO J. 16, 3705-3714 (1997).</li><li>Harrington, L. A., Greider, C. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Telomerase+primer+specificity+and+chromosome+healing.">Telomerase primer specificity and chromosome healing.</a> Nature. 353, 451-454 (1991).</li><li>Jacob, N. K., Skopp, R., Price, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=G-overhang+dynamics+at+Tetrahymena+telomeres.">G-overhang dynamics at Tetrahymena telomeres.</a> Embo J. 20, 4299-4308 (2001).</li><li>Barry, J. D., McCulloch, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antigenic+variation+in+trypanosomes:+enhanced+phenotypic+variation+in+a+eukaryotic+parasite.">Antigenic variation in trypanosomes: enhanced phenotypic variation in a eukaryotic parasite.</a> Adv Parasitol. 49, 1-70 (2001).</li><li>Yang, X., Figueiredo, L. M., Espinal, A., Okubo, E., Li, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=RAP1+is+essential+for+silencing+telomeric+variant+surface+glycoprotein+genes+in+Trypanosoma+brucei.">RAP1 is essential for silencing telomeric variant surface glycoprotein genes in Trypanosoma brucei.</a> Cell. 137, 99-109 (2009).</li></ol>

布氏锥虫引起睡在人类的疾病。这种疾病，如果不及时治疗，必然是致命的 T 。 布氏在哺乳动物宿主细胞进行定期抗原变异，以逃避宿主的免疫攻击7。因此，它是非常难以消除T。布氏一旦感染细胞的建立。最近，我们已经证明， 端粒 T的表达调控发挥了重要作用布氏表面抗原基因8。因此，重要的是进一步了解在 T端粒维持端粒的?...

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examination of the telomere g-overhang structure in trypanosoma brucei

端粒的G -悬结构已被确定在许多真核生物，包括酵母，脊椎动物和布氏锥虫。它作为基板从头端粒DNA合成的端粒酶，因此，重要的端粒维持 T 。 布氏是一种原生动物寄生虫引起的睡眠疾病在人类和牛的那加那病。一旦被感染的哺乳动物主机， T.定期交换机布氏细胞的表面抗原，以逃避宿主的免疫攻击。我们最近表明的T.布氏端粒的结构起着至关重要的作用，在调控的表面抗原基因的表达，这是 T的关键布氏的发病机制。然而，T.布氏端粒的结构还没有被广泛研究，由于这个专门的结构分析方法的局限性。我们现在已经成功地通过在凝胶杂交和连接介导的引物延长端粒G -悬结构， 在 T核苷酸测定端粒终端适配器结扎方法检查方法的原生布氏细胞。在这里，我们将详细描述了协议，并比较其不同的优势和局限。

Watch this Scientific Journal Video about Examination of the Telomere G-overhang Structure in Trypanosoma brucei at JoVE.com

Examination of the Telomere G-overhang Structure in Trypanosoma brucei

端粒是染色体的稳定至关重要，并介导的端粒酶的端粒维持端粒G -悬结构是必不可少的。最近，我们通过两种方法检测端粒G -悬结构<em&gt;锥虫布氏</em&gt;，这是原生 - 凝胶杂交和连接介导的引物延伸，这将是描述。

审核端粒G -悬结构锥虫布氏</em

The telomere G-overhang structure has been identified in many eukaryotes including yeast, vertebrates, and Trypanosoma brucei. It serves as the substrate ...

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