SV40早期启动子中原核增强子样序列的初步研究  

A Primary Study on Prokaryotic Enhancer-like Element in SV40 Early Promoter

作　　者：李植峰[1] 高丽杰[1] 施庆国[1] 曹韫旭[2] 孙树秦[1] 陆德如[2] 

机构地区：[1]解放军北京军医学院生物化学教研室,北京100071 [2]第二军医大学分子遗传学研究所,上海200433

出　　处：《科技通报》2002年第1期20-23,共4页Bulletin of Science and Technology

摘　　要：萤火虫荧光素酶基因 c DNA真核表达载体 p GL2 可在原核细胞中高效表达 .该载体中含有 SV4 0早期启动子及荧光素酶基因 c DNA 5’端总长共 75 8bp的 DNA碱基序列 ,经计算机分析发现 ,在 SV4 0早期启动子中含有 SV4 0基因组 Hind B片断中一段长为 4 9bp的 DNA序列 ,且荧光素酶基因 c DNA编码区的 5’端含有一个原核细胞基因表达调控区相似序列 .将 p GL2 中的荧光素酶基因 c DNA(L uc)克隆入真核表达载体 p ED中 ,构建成 p ED- Luc.该载体经表达测定后 ,只在真核细胞中表达 ,在原核细胞中不表达 .这说明荧光素酶基因 c DNA编码区的 5’端的原核细胞基因表达调控区相似序列不具表达有活性的荧光素酶的功能 .经 DNA序列对比研究 ,认为 p GL2 含有的 SV4 0基因组 Hind B片段 5 12 3bp～ 5 171bp长 4 9bp的 DNA序列可能就是 Hind B片段的原核增强子功能的决定序列 .The eukaryotic expressing vector pGL 2 of luciferase gene cDNA can express efficiently in prokaryotic cells. Using the PC/GENE procedure to analyse the 758bp length DNA sequence including SV40 early promoter and the 5-end of luciferase gene cDNA, a 49bp length DNA sequence belong to the SV40 HindⅢ B fragment was found in the SV40 early promoter of pGL 2, and there is a prokaryotic gene expression control-like element in 5-end of luciferase gene cDNA.The luciferase gene cDNA(Luc) of pGL 2 was clonded into eukaryotic expressing vector pED to construct plasmid pED-Luc,it can express luciferase only in eukaryotic cells,but not in prokaryotic cells.It showed that the prokaryotic gene expression control-like element in 5-end of luciferase gene cDNA is inable to express a activity luciferase.The result of contrasting DNA sequence showed that the 49bp length DNA sequence belong to the SV40 HindⅢ B fragment 5123bp-5171bp could be the decisive sequence to the prokaryotic enhancer function of the SV40 HindⅢ B fragment.

关 键 词：SV40早期启动子 荧光素酶基因 原核增强子 

分 类 号：Q78[生物学—分子生物学]