[생화학] HBV 유전자 복제에 작용하는 core protein의 규명

목차

I. 서론
1. Hepatitis B virus 및 hepadnavirus family
2. Hepatitis B virus 유전자 구조
3. Infection cycle
4. Encapsidation
5. Replication
5.1 Replication mechanism
5.2 HBV genome의 replication intermediate

II. 실험재료 및 방법
1. 실험재료
2. 실험방법
2.1 Plasmids
2.1.1 HBV expression plasmids
2.1.2 WHV expression plasmids
2.1.3 Chimeric core expression plasmids
2.2 세포 배양 및 transfection
2.2.1 세포배양
2.2.2 Transfection
2.3 Core particle의 분리 및 viral DNA추출
2.4 Southern blot analysis
2.4.1 Gel electrophoresis
2.4.2 Denaturation
2.4.3 Neutralization
2.4.4 Nylon membrane으로 transfer
2.4.5 Probe 제조
2.4.6 Hybridization
2.4.7 Autoradiography
2.5 RNase protection assay
2.5.1 RPA probes의 Preparation
2.5.2 Hybridization
2.5.3 RNase digestion
2.5.4 Gel electrophoresis
2.5.5 Autoradiography

III. 실험결과
1. 연구 목적
2. Plasmid construction
3. HBV genome replication에서의 WHV core에 의한 Complementation
4. HBV genome replication에서의 WHV polymerase에 의한 Complementation
5. Chimeric core proteins의 Replicative intermediate 생성 경향 분석
6. Chimeric core proteins의 Replicative intermediate 생성 경향 분석
7. Chimeric core proteins의 encapsidation efficiency 조사

IV. 고찰

V. 참고문헌
영문요약

본문내용

Hepatitis B virus (HBV)는 hepadnavirus의 일종으로서 종 특이성과 조직 특이성이 있는 바이러스로서 DNA를 genome을 갖는 바이러스로서 만성적인 간 질환과 hepatocellular carcinoma를 일으키는 주원인으로 알려져 있다. Hepadnavirus family에는 woodchuck hepatitis virus (WHV), ground squirrel hepatitis virus, duck hepatitis B virus (DHBV) 등이 있다. 이중 WHV는 woodchuck이라는 포유 동물을 숙주로 하는 바이러스로 HBV와의 nucleotide sequence homology는 78 %이다. Hepadnavirus는 RNA intermediate인 pregenomic RNA의 reverse transcription에 의해 replication된다. Replication의 첫 단계는 pregenomic RNA가 core particle내로 encapsidation되는 것이다. Encapsidation은 structural protein인 core와 polymerase 그리고 pregenomic RNA의 interaction에 의해 이루어진다. 먼저 polymerase는 pregenomic RNA의 5´ end에 있는 stem-loop 구조인 ε 에 결합하게 되며 이 polymerase-pregenomic RNA complex를 core protein이 인식해 encapsidation이 이루어진다. 이렇게 encapsidation이 일어나고 나면 capsid 내에서 pregenomic RNA의 reverse transcription으로 DNA synthesis가 이루어진다.

참고 자료

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