双位点核酶抑制细胞内HBV表达的图像分析

目的：利用ＬＥＩＣＡＱ５００ＭＣ图像分析系统观察针对ＨＢＶＣ区双位点核酶对细胞质中Ｃ区基因表达的抑制作用。方法：采用亚克隆技术，从ｐＧＥＭＲｚ１２３（含针对ＨＢＶＣ区双位点核酶）上切下双位点核酶的片段，定向克隆于真核表达载体ｐＢＢＳ２１２中，利用Ｌｉｐｏｆｅｃ－ｔａｍｉｎｅ介导，将重组质粒ｐＢＢＳ２１２－Ｒｚ转染入２２１５细胞中，采用原位杂交观察核酶的表达，采用免疫组化、图像分析法分析ＨＢｃＡｇ的表达。结果：筛选２周后，原位杂交证实可表达针对ＨＢＶＣ区双位点核酶。免疫组化及图像分析证实该核酶可抑制Ｃ区基因表达。结论：应用图像分析法可定量地观察到核酶对Ｃ区基因表达有阻断作用，抑制ＨＢｃＡｇ的合成。

Image analysis of inhibition of HBcAg by twounit ribozymegene against hepatitis B virus

By using LEICA Q 500MC image analysis system, to observe the inhibition of HBcAg expressed in the 2.2.5 cell tranafected by twonuit ribozyme.Methcds:With the subclone technique, the twounit ribozyme gene cut from pGEM-Rz123 was ligated into the eukaryotic expression vectorpBBS212. The recombinant pBBS212-Rz plasmids were transfected into the 2.2.15 cells mediated by Lipofectamine. The ribozyme was dectected by dot blot. The c antigen of HBV was detected by immunohistochemical technique and image analysis system.Result: By using immunohistochemical technique and image analysis system,it revealed that the level of intracelluar HBcAg expression went down. Conclusion:All the results strongly indicate that the twounit ribozyme may inhibit the HBV expression in the cell.