核酶对2.2.15细胞中HBeAg表达抑制作用的初步观察

INHIBITION OF HBeAg EXPRESSION IN 2.2.15 CELL BY TWO-LOCUS RIBOZYME

目的：观察针对ＨＢＶ Ｃ 区双位点核酶对２ ．２ ．１５ 细胞中ＨＢｅＡｇ 表达的抑制作用。方法：利用亚克隆技术，从质粒ｐ ＧＥＭ － Ｒｚ１２３ 切下ＥｃｏＲ Ｉ－ Ｂａｍ Ｈ Ｉ 片段克隆于真核表达质粒ｐＢＢＳ２１２ 中，将该质粒转染２ ．２ ．１５ 细胞，用ＥＬＩＳＡ 及免疫组化方法观察该核酶对ＨＢｅＡｇ 合成的抑制作用。结果：细胞中表达出针对ＨＢＶ Ｃ 区核酶，该核酶对ＨＢｅＡｇ 的抑制率达４８ ．６ ％ 。结论：该双位点核酶可能通过针对ＨＢＶ Ｃ 区基因的剪切作用，阻断Ｃ

jective:To observe the inhibition of HBeAg expression in the 2.2.15 cell transfected by two-locus ribozyme against HBV.Methods:The two-locus ribozyme gene, cut from plasmid pGEM-Rz123, was cloned into the eukaryotic expression vector pBBS212. The recombinant plasmid pBBS212-Rz was transfected into 2.2.15 cell using lipofectamine. The supernate antigen of HBV was detected by ELISA and immunohistochemistry to clarify the inhibition of HBeAg.Results:Ribozyme could be expressed in 2.2.15 cell by in-situ hybridization.Meanwhile,this two-locus ribozyme could reduce the level of HBeAg expression in 2.2.15 cell by up to 48.6%.Conclusions:The two-locus ribozyme can inhibit the expression of HBeAg in transfected 2.2.15 cell by cutting the relative HBV gene.