STUDY ON STRUCTURE AND FUNCTION OF PREINTEGRATION COMPLEX OF HUMAN IMMUNODEFICIENCY VIRUS TYPE 1

为了检测基于具有病毒中央多嘌呤段（cPPT）的人类免疫缺陷病毒1型（HIV）的载体是否比不具有cPPT的载体更有效地表达转基因，我们构建了一系列携带Neo '基因作为稳定标记的载体，该基因由内部合成的胸苷激酶启动子驱动。插入282bp或178bp长的cPPT导致产生的载体颗粒的量增加约3 - 8倍。插入较长的cPPT。导致转导效率降低约八倍，而插入较短的导致转导效率增加约两倍，与插入物的取向无关。DNA-PCR结果显示，逆转录和核内导入不受影响。RT-PCR检测表明，282bp-cPPT基因在载体中的整合受到了破坏。综上所述，我们得出结论，282bp-cPPT的插入减少了0418抗性菌落的数量，而不影响载体DNA的核输入。我们假设282bp-cPPT的插入导致导入后事件的损害，例如整合。

To examine whether vectors based on human immunodeficiency virus type 1 (HIV) with viral central polypurine tract (cPPT) express transgenes more efficiently than those without cPPT, we constructed a series of vectors carrying a Neo'gene as a stable marker, which is driven by an internal synthetic thymidine kinase promoter. Insertion of cPPT, either 282bp or 178bp long, resulted in about 3-8-fold increase of the amount of produced vector particles. Insertion of the longer cPPT. resulted in decrease in transduction efficiency in about eight-fold, whereas insertion of the shorter resulted in increase in transduction efficiency in about two-fold independently of the orientation of the insert. Moreover, DNA-PCR showed that reverse transcription and nuclear import were cnot affected. Alu-PCR assay showed that integration was impaired in the vectors carrying the 282bp-cPPT. Taking these together, we concluded that insertion of the 282bp-cPPT decreased the number of 0418-resistant colonies without affecting nuclear import of vector DNA. We suppose that insertion of the 282bp-cPPT resulted in impairment of events after import, such as integration.