BIV－LTR中存在负调控区

NEGATIVE REGULATORY ELEMENTS IN BIV LTR

为研究ＢＩＶ－ＬＴＲ调控序列的功能，将ＬＴＲ部分ＤＮＡ序列缺失，与荧光素酶基因相连构建质粒ｐＤ－３１９－Ｌｕｃ，ｐＤ－２６１－Ｌｕｃ，ｐＤ－１８１－Ｌｕｃ，ｐＤ－２－Ｌｕｃ，ｐＤ＋３２－Ｌｕｃ，ｐＤ－３１９－２６１－Ｌｕｃ，ｐＤ－１８１＋３２－Ｌｕｃ．通过磷酸钙共沉淀法将上述质粒及ｐＢＬＴＲ－Ｌｕｃ分别转染小牛肺细胞（ＦＢＬ），检测转染细胞裂解物荧光素酶活性，比较各质粒所克隆的ＢＩＶ－ＬＴＲ表达水平，发现－２６１位上游区存在负调控区，Ｒ区存在抑制ＬＴＲ表达的序列．

To Study the functions of BIV LTR regulatory; elements, the LTR was partially deleted and linked to luciferase gene, hence,we constructed a series of plasmids: pD-319-Luc,pD-261-Luc,pD-181-Luc,pD-2-Luc,PD+32-Luc, pD-319-261-Luc and pD-181+32-Luc. Using the phosphate-calcium co-transfection technique, these plasmids and pBLTR-Luc was transfected into Fetal bovine lung cell (FBL),respectively.Through the detection of luciferase activities in the lysised debries of the transfected cells,we compared the expression levels of each cloned D BIV LTR,and found that DNA sequence upstream of-261 site can repress the expression of LTR,meanwhile,there is a negative regulatory region in the R region of LTR as well.