ANALYSIS OF HIV MESSENGER RNA STRUCTURE

人类免疫缺陷病毒(HIV)感染通过以下方式显示 几个实验室使用一套复杂的RNA合成和 处理事件以控制病毒表达。按顺序 确定是否存在以前未发送过的消息 我们已经开始直接分析HIV的cdna拷贝。 经Southern杂交和DNA序列分析。 从分离的rna中获得了lambda gt10的cdna文库。 来自感染艾滋病毒的H9细胞。从这里产生的大量噬菌体 文库被发现与病毒序列杂交。共有19个 这些噬菌体是经噬菌体纯化和Southern分析的 利用标记的寡核苷酸或限制性内切酶进行杂交 来自艾滋病毒基因组不同位置的片段。这个 具体区域包括：(A)TAT的5‘端：(B)TRS的5’端；(C) 5‘-环境部分(不包括TAT和TRS区域)；(D)POL/SOR 区域。具有以下杂交模式的克隆已被 隔离：I类，A+B-C-D+；II类，A-B-C+D+；III类，A-B-C- D+。我们已经纯化了其中的几个噬菌体，并进行了亚克隆 他们将DNA插入pIBI载体。其中几个噬菌体具有 经Southern印迹杂交鉴定。我们是 目前正在对这些DNA片段进行测序以确定它们是否 代表新的HIV信息，因此可能揭示新的外显子或 拼接位置。

Human immunodeficiency virus (HIV) infection has been shown by several laboratories to use a complex set of RNA synthesis and processing events in order to control viral expression. In order to determine if messages exist that have not been previously characterized, we have begun to directly analyze cDNA copies of HIV messages by Southern hybridization and DNA sequence analysis. A cDNA library in lambda gt10 has been prepared from RNA isolated from HIV-infected H9 cells. A large number of phages from this library were found to hybridize to viral sequences. A total of 19 such phages were plaque-purified an' analyzed by Southern hybridization using labeled oligonucleotides or restriction fragments derived from various locations on the HIV genome. The specific region include: (A) 5'-end of tat: (B) 5'-end of trs; (C) 5'-portion of env (excluding tat and trs region); (D) pol/sor region. Clones with the following hybridization pattern have been isolated: class I, A+B-C-D+; class II, A-B-C+D+; class III, A-B-C- D+. We have purified several of these phages and have subcloned their insert DNA into pIBI vectors. Several of these phages have been characterized by Southern blot hybridization. We are currently sequencing these DNA segments to determine if they represent novel HIV messages and, thus, might reveal new exons or splice sites.