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近年兴起的反义技术为开发抗病毒药物带来了新的希望,其中反义寡聚脱氧核苷酸(antisense oligodeoxynucleotidc,as ODN)抗人乙型肝炎病毒(HBV)的研究是其热点之一。通过互补于HBV基因或其mRNA的人工合成的asODN可达到抗HBV的目的。为了增强抗病毒效果,本文设计了针对HBV2.1KbmRNA起始区、表面抗原基因起始区、e抗原基因起始区的三个硫代磷酸反义寡聚脱氧核苷酸片段。根据固相亚磷酸酰胺合成法,利用DNA合成仪,合成了均为18个碱基长度的三个as ODN片段。经过从固相载体上的切下、去保护基和变性聚丙稀酰胺凝胶电泳的纯化与鉴定,采用浸泡法最终回收到了所需的as ODN片段并测定了其DNA含量。
The antisense technology that has emerged in recent years has brought new hope for the development of antiviral drugs. Among them, antisense oligodeoxynucleotidc (as ODN) is one of the hot spots in its study of anti-human hepatitis B virus (HBV) . The purpose of anti-HBV is achieved by synthetic asODN complementary to the HBV gene or its mRNA. In order to enhance the antiviral effect, three phosphorothioate antisense oligodeoxynucleotide fragments targeting HBV2.1Kb mRNA initiation region, surface antigen gene initiation region and e antigen gene initiation region were designed. According to the solid-phase phosphite synthesis method, three as ODN fragments each having an 18-base length were synthesized by using a DNA synthesizer. After purification and identification from excised, deprotected and denatured polyacrylamide gel electrophoresis on a solid support, the desired as ODN fragment was finally recovered by immersion and the DNA content was determined.