单纯疱疹病毒DNA探针制备的研究

The Preparation of HSV DNA Probe

本文从克隆化的HSV-1 EcoR Ⅰ-H片段重组菌株分离目的基因EcoR Ⅰ-H片段（13,5kb）。用缺口翻译法制备32P标记探针,通过点杂交试验检测HSV-1,HSV-2,腺病毒3型（ADV3）,痘苗病毒天坛株（VV）,pBR322,Vero cell DNA及含EcoR Ⅰ-H片段的重组质粒DNA。结果表明,该探针只与HSV-1、HSV-2 DNA杂交,而不与ADV3,VV,Vero-cellDNA杂交。且可检测出至少5pg的同源序列DNA。表明此探针具有良好的特异性和敏感性。

EcoR 1-Hfragment (13,5kb) was selected from cloned HSV-1 EcoR I-H fragment. ^(32)P-labeled DNA probe was prepared by nick translation. DNAs obtained from HSV-1,HSV-2, ADV_3. VV,_PBR_(322),EcoR I-H fragment and Vero cell were detected by dot hybridization assay with this Probe. The results showed that the probe hybridizated only with HSV-1 and HSV-2 DNA, but not with ADV_3, VV,Vero cell DNA. The probe can detect 5pg of DNA sequence homology, Therefore,the probe was thought to be specific and sensitive.