| Telomerase Repeated Amplification Protocol (TRAP)
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Author:
Date:
2015-11-20
[Abstract] Telomeres are found at the end of eukaryotic linear chromosomes, and proteins that bind to telomeres protect DNA from being recognized as double-strand breaks thus preventing end-to-end fusions (Griffith et al., 1999). However, due to the end replication problem and other factors such as oxidative damage, the limited life span of cultured cells (Hayflick limit) results in progressive shortening of these protective structures (Hayflick and Moorhead, 1961; Olovnikov, 1973). The ribonucleoprotein enzyme complex telomerase- consisting of a protein catalytic component hTERT and a functional RNA component hTR or hTERC- counteracts telomere shortening by adding telomeric repeats to the end of chromosomes in ~90% of primary human tumors and in some transiently ...
[摘要] 端粒存在于真核线性染色体的末端，并且结合端粒的蛋白质保护DNA不被识别为双链断裂，从而防止端对端融合（Griffith等人，1999） 。然而，由于末端复制问题和其他因素如氧化损伤，培养细胞的有限寿命（Hayflick极限）导致这些保护结构的逐渐缩短（Hayflick和Moorhead，1961; Olovnikov，1973）。由蛋白质催化组分hTERT和功能性RNA组分hTR 或组成的核糖核蛋白酶复合物端粒酶端粒酶通过添加端粒重复来抵消端粒缩短到约90％的原发性人类肿瘤和一些短暂增殖的干样细胞中染色体的末端（Shay和Wright，1996; Shay和Wright，2001）。这导致细胞的连续增殖，这是癌症的标志。因此，端粒生物学在衰老，癌症进展/转移以及靶向癌症治疗中具有中心作用。端粒生物学中常用的方法如端粒限制性片段（TRF）（Mender和Shay， 2015b），端粒重复扩增方案（TRAP）和端粒功能障碍诱导Foci（TIF）分析（Mender和Shay，2015a）。在这个详细的协议中，我们描述端粒重复扩增协议（TRAP）。 ...
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| Introduction and Sequencing of Patient-isolated HBV RT Sequences into the HBV 1.2-mer Replicon
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Author:
Date:
2015-04-20
[Abstract] Antiviral agents for the suppression of hepatitis B virus (HBV) have been used for treating chronic hepatitis B. However, the emergence of drug-resistant HBV is still a major problem for antiviral treatment. To identify and characterize the drug-resistant HBV, the construction of HBV replicon and in vitro drug susceptibility assay are essential. Here we describe the experimental methods to study drug-resistant HBV.
[摘要] 用于抑制乙型肝炎病毒（HBV）的抗病毒剂已经用于治疗慢性乙型肝炎。然而，耐药性HBV的出现仍然是抗病毒治疗的主要问题。 为了鉴定和表征耐药性HBV，构建HBV复制子和体外药物敏感性测定是必需的。 在这里我们描述研究耐药性HBV的实验方法。
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