FEN 1基因反义阻断细胞株(FL FEN 1^-)的建立  

Establishment of Cell Line with the FEN 1 Gene Blocked by Its Antisense

作　　者：石斌山[1] 余应年[1] 蔡朱男[1] 沈炳辉[2] 

机构地区：[1]浙江大学医学院病理生理教研室,杭州310031 [2]CityofHopeNationalMedicalCenterandBeckmanResearchInstitute

出　　处：《生物化学与生物物理学报》2000年第1期87-89,共3页

基　　金：国家自然科学基金

摘　　要：ＦＥＮ１ 是一种结构特异性核酸酶， 它在ＤＮＡ 复制和修复过程中都起着重要的作用。将ＦＥＮ１ 基因的ＮｃｏＩＢａｍ ＨＩ片段反向克隆到哺乳动物细胞重组表达质粒ｐＭＡＭｎｅｏＡｍｐ－ 中， 得到ＦＥＮ１ 反义表达质粒ｐＭＡＭｎｅｏＡｍｐ－ ＦＮＢ－ ， 并转染ＦＬ细胞， 经Ｇ４１８ 筛选后， 获得ＦＥＮ１ 基因表达被阻断的哺乳动物细胞ＦＬＦＥＮ１－ 。对细胞生长曲线的测定发现，ＦＬＦＥＮ１－ 在地塞米松的诱导下， 细胞生长速度明显下降， 细胞倍增时间ＴＤ 为３ ．０３ｄ， 而ＦＬ 和仅含有空载体的ＦＬＭ 细胞在地塞米松存在下的细胞倍增时间ＴＤ 分别为２．０３ ｄ 和２ ．２２ ｄ， 不经地塞米松诱导的ＦＬＦＥＮ１－ 细胞的ＴＤ 为２ ．３８ ｄ 。FEN 1 is a structure specific endo/exonuclease, which is involved in the process of both DNA duplication and DNA repair. In this work a mammalian expression vector expressing antisense FEN 1 gene fragment pMAMneoAmp -FNB - was constructed, after cloning the Nco I Bam HI fragment of FEN 1 gene into the mammalian expression vector pMAMneoAmp - in antisense orientation. After FL cell was transfected with pMAMneoAmp -FNB - and selected by G418, the FL FEN 1 - cell line, in which the FEN 1 gene expression was blocked, was established. It was found that the growth of FL FEN 1 - was decreased upon the induction with dexamethasone and its T D was 3.03 d, while the T D of controls FL and FL M induced with dexamethasone was 2.03 and 2.22 d, respectively, and the T D of the FL FEN 1 - cell without dexamethasone was 2.38 d.

关 键 词：结构特异性核酸酶FEN-1 反义核酸技术 FL细胞 

分 类 号：Q782[生物学—分子生物学]