HIV REPORTER CELL LINE FOR SCREENING ANTI-SENSE AGENTS

一个分泌型碱性磷酸酶（SPAP）报告基因将被取代 HIV前病毒DNA克隆中env基因的一部分。 表达 报告基因将由病毒顺式完全互补物调节 和反式作用控制元件，使得细胞外的 碱性磷酸酶将作为病毒gag、pol和env的指示剂 受病毒编码达特、rev和nef正常调节的区域 基因产物 将用该构建体转染H9细胞系 SPAP、p24和逆转录酶的瞬时表达 表征了 用SPAP作为标记物检测 将通过监测三种药物的效果来评估抗病毒活性 靶向HIV关键位点的反义寡核苷酸（ODNs） 转染的H9细胞中的基因组。

A secretable alkaline phosphatase (SPAP) reporter gene will be substituted for a portion of the env gene in an HIV proviral DNA clone. Expression of the reporter gene will be regulated by a complete complement of viral cis and trans acting control elements, such that the level of extracellular alkaline phosphatase will serve as an indicator of viral gag, pol, and env regions subject to normal regulation by the viral coded tat, rev, and nef gene products. The H9 cell line will be transfected with this construct and the transient expression of SPAP, p24, and reverse transcriptase characterized. The feasibility of using the SPAP as a marker to detect antiviral activity will be assessed by monitoring the effect of three antisense oligonucleotides (ODNs) targeted to critical sites on the HIV genome in the transfected H9 cells.