论文部分内容阅读
目的:构建针对尿激酶型纤溶酶原激活剂(uPA)mRNA的siRNA的表达载体,并观察其对转染细胞中的uPA表达的抑制作用.方法:化学合成用于产生针对uPA的发卡状RNA的寡核苷酸,各64个碱基,退火后插入线性化的pSU PER载体的H1启动子下游.重组载体经限制性酶切和测序.把构建的载体转染乳腺癌细胞系MDA MB231,观察其对uPA的干扰作用.结果:限制性酶切和序列测定表明成功构建了可以表达针对uPA的发卡状RNA表达载体,该载体表达的发卡状RNA能够有效的下调uPA的表达.结论:成功的构建了针对uPA的发卡状RNA表达载体,为进一步研究uPA的功能和肿瘤的基因治疗打下基础.
OBJECTIVE: To construct a siRNA expression vector targeting urokinase-type plasminogen activator (uPA) mRNA and to observe its inhibitory effect on uPA expression in transfected cells.Methods: The chemical synthesis was used to generate hairpin RNA oligonucleotides, each 64 bases, annealed downstream of the H1 promoter of the linearized pSU PER vector.The recombinant vector was restriction digested and sequenced.The constructed vector was transfected into the breast cancer cell line MDA MB231 , And observed its interference with uPA.Results: Restriction endonuclease digestion and sequencing showed that the hairpin RNA expression vector expressing uPA gene was successfully constructed, and the hairpin RNA expressed by this vector can effectively down-regulate uPA expression.Conclusion: The successful construction of hairpin RNA expression vector targeting uPA laid the foundation for further study on the function of uPA and gene therapy of tumor.