TEST FOR HIV-I REV PROTEIN USING O-BETA SOLID TECHNOLOGY

描述：(改编自申请人摘要)申请人建议 建立HIV-1 Rev蛋白的临床分析方法。技术 是基于两种相对较新的技术的组合。首先是 到目前为止已经识别出强结合核苷酸的SELEX技术 Rev蛋白的序列。二是Q-的特定扩增 核糖核酸酶，核糖核酸酶。这个 这个想法是制造两个RNA适配子，它将以这样的方式与REV结合 末端并列，用于T4RNA连接酶的连接。一旦这一次 完成后，连接的RNA可以被Q-beta特异性地扩增 复制酶。扩增产物可作为REV的定量 蛋白。为了开发这样一个系统，申请人提议 构建两个REV蛋白结合和For的重组RNA适配子 Q-β复制酶模板。然后找出适配子的条件 在它们自我退火之前，最大限度地退火到REV。这将是 然后在连接和扩增中进行检测。如果这成功了， 检测试剂盒中的方法将使用临床样本进行测试。

DESCRIPTION: (Adapted from Applicant's Abstract) The applicant proposes to develop a clinical analytical method for HIV-1 Rev protein. The technology is based on the combination of two relatively new technologies. First is the SELEX technology which has so far identified strong binding nucleotide sequences to Rev protein. The second is the specific amplification of Q- beta replicase of RNA containing recognized sequences by this enzyme. The idea is to make two RNA apttamers that will bind to Rev in such a way that the ends are juxtapositioned for the ligation by T4 RNA ligase. Once this is done, the ligated RNA can be amplified specifically by Q-beta replicase. The amplified product would be used as quantitation of Rev protein. For the development of such a system, the applicant proposes to construct two recombinant RNA aptamers for Rev protein binding and for the template of Q-beta replicase. Then to find conditions the aptamers anneal maximally to Rev before they anneal to themselves. This will be followed by testing in ligation and amplification. If this is successful, the methods in test-kits will be tested using clinical samples.