摘要
利用 Ca Cl2 法成功地将寡核苷酸导入大肠杆菌 JM1 0 9细胞。质粒 p BR32 2转化结果表明 ,寡核苷酸片段 5′- AGCGGGAATAAGGGAA- 3′在转化前 (或后 )与靶序列结合均能抑制 β-内酰胺酶基因的表达 ,细菌生长受到抑制。体外聚丙烯酰胺凝胶电泳结果表明 ,该片段能与靶序列形成三链结构。这些结果表明多聚嘌呤寡核苷酸是通过与靶序列形成三链 DNA来抑制β-内酰胺酶基因的表达。
The oligonucleotides have been introduced into Escherichia coli JM109 by the CaCl 2 procedure. The results of transformation showed that the gene expression of Amp r and the growth of the transformed bacteria were inhibited by the purine rich oligonucleotide 5′ AGCGGGAATAAGGGAA 3′ before or after the transformation of plasmid pBR322. Compared with purine rich oligonucleotide, the pyrimidine rich oligonucleotide hardly showed any inhibition to the gene expression of Amp r and the growth of the transformed bacteria. The result of gel electrophoresis also showed that the purine rich oligonucleotide could easily form a oligonucleotde directed triple helix with the targeted sequence of plasmid pBR322. These results supported that the gene expression of Amp r was inhibited by oligonucleotide via the oligonucleotide directed triple helix formation.
出处
《华东理工大学学报（自然科学版）》
CAS
CSCD
北大核心
2000年第6期608-611,共4页
Journal of East China University of Science and Technology
基金
国家自然科学基金!(396 80 0 32 )
国家科委医药技术创新博士项目
霍英东基金资助
