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： 开发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结合。一是 到目前为止已经鉴定出强结合核苷酸SELEX技术 Rev蛋白的序列。二是Q-的特异性放大作用， RNA的β复制酶，含有这种酶识别的序列。的 我们的想法是制造两个RNA适体，它们将以这样一种方式与Rev结合， 将末端并置以通过T4 RNA连接酶进行连接。一旦这个 完成后，连接的RNA可以通过Q-β特异性扩增， 复制酶扩增产物将用作Rev 蛋白为了开发这样的系统，申请人建议 构建两个重组RNA适体，用于Rev蛋白结合和 Q-β复制酶的模板为了找到适体 在它们退火到自身之前，最大程度地退火到Rev。这将是 随后进行连接和扩增测试。如果成功的话， 将使用临床样本对测试包中的方法进行测试。

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.