TrkAsiRNA表达载体构建及对TrkA基因表达的抑制

Construction of siRNA expression vector of trkA and its inhibition on the expression of trkA

目的 构建具有特异阻断TrkA基因功能的siRNA表达系统 ,为前列腺癌基因治疗提供新的方法。方法 根据Genbank提供的TrkA基因mRNA序列 ,应用设计软件设计特异性的短链寡核苷酸 ,化学合成后经退火形成双链DNA片段 ,克隆到pSlincerTM2 1 U6载体中 ,用BamHⅠ和HindⅢ双酶切和序列测定对重组体进行鉴定 ,最后将构建的表达载体转染前列腺癌细胞株 ,观察对TrkA基因表达的影响。结果 酶切和序列测定表明 ,成功构建了siRNA表达载体 ,能够抑制细胞内TrkA基因表达。结论 本研究构建的siRNA表达载体 ,具有阻断TrkA基因的表达的功能 ,为前列腺癌基因治疗提供新的有效的方法。

Objective To construct the specific siRNA expression vector that can block TrkA gene function, which provides a new approach to the prevention of prostate cancer. Methods Specific short chain oligonucletide was designed by using the siRNA software according to the mRNA sequence provided by genebank, the double chain DNA sequenct was gained through annealing after chemosynthesis and was inserted to psilencer siRNA expression vector, the recombinant expression vector was evaluated by using enzyme cutting and sequencing. At last, the constructed TrkA gene expression was observed. Results Identifcation by enzyme cutting and sequencing showed that the expression vector was constructed successfully, and could inibit the gene expression effiectively. Conclusion The constructed siRNA expression vector can block the TrkA gene expression, it will enable us to develop an approach to cure prostate cancer.