盐芥miRNA393沉默载体的构建

Construction of miRNA393 Silenced Vector in Thellungiella salsuginea

将与内源mRNA编码区同源的小片段外源双链RNA导入细胞中,通过一系列细胞内反应引发细胞中相应mRNA降解,可导致特定基因表达沉默。通过PCR扩出与miRNA399互补的IPS,并改造成与miRNA393互补的IPS′,测序正确后酶切连入3301H载体转化盐芥。

When guides small exogenous dsRNAs to cells that are homologic to their endogenesis RNA＇s protein coding region ,they can lead specific gene to silence by series of reactions which bring in-cell＇s corresponding mRNA to degrade. The gene IPS which complementary to miRNA399 sequence was firstly received and it was reconstructed to IPS＇ that contains a region complementary to miRNA393. After making sure the sequce right, they connected the aid gene with the 3301H empty vector.