核酶的体外合成及其对鸭乙型肝炎病毒S基因体外转录物的定点切割  

HAMMERHEAD RIBOZYME-MEDIATED CLEAVAGE OF IN VITRO TRANSCRIPTS OF DUCK HEPATITIS B VIRUS SURFACE CENE

作　　者：汤华[1] 闻玉梅[1] 何丽芳 

机构地区：[1]上海医科大学卫生部医学分子病毒学重点实验室

出　　处：《病毒学报》1994年第4期307-310,共4页Chinese Journal of Virology

基　　金：国家教委博士点基金

摘　　要：选择鸭乙型肝炎病毒作为核酶的靶病毒，设计针对病毒前基因组Ｓ基因区第１２８８位和１４６９位“锤头状”核酶序列（ＲＺ１和ＲＺ２）。经体外转录及做核酶切割，ＲＺ１和ＲＺ２均有切割作用，以ＲＺ１作用更强。鉴于核酶在相当于鸭体温（４２℃）有切割作用，因此有可能在鸭体内研究其切割作用。ccording to the “hammerhead structure”described by Symons, we designed two ribozymesaimed at the 1228nt and 1467 nt of the pregenonme RNA of duck hepatitis B virus.cDNAs of ribozyme RZ1,RZ2 were synthesized by an automated DNA synthesizer,ligated to pGEM4zplasmid and transcribed in vitro with T7 RNA polymerase for preparation of ribozymes.pGDT1plasmid was constructed by insertion of SmaI/AccI fragment (1179-1585 nt)of DHBVDNA into the saim sites of pGDT1 digested with AccI was transcribed by T7 RNA polymerase and radiolabeled with 32p-UTP,and served as the substrate.One pmol ofribozyme RZ1 or RZ2 and one pmol of substrate were incubated at 42℃ 2 hours for cleavage. Data indicate that both ribozymes could cleave its substrate, while RZ1 was moreeffective. Since 42℃ is the body temperature of ducks, it is possible that these ribozymes can be used for in vivo studies of their cleavage effects in the DHBV infected animal model.

关 键 词：核酶 鸭乙型肝炎病毒 切割 合成 S基因 转录物 

分 类 号：R373.21[医药卫生—病原生物学] Q939.403[医药卫生—基础医学]