Investigation of HDV antigen induced genes by FISH-PCR

• Main Authors: Jui-Pin, 張瑞玶
• Other Authors: 許國堂
• Format: Others
• Language: zh-TW
• Published: 2005
• Online Access: http://ndltd.ncl.edu.tw/handle/07961534503583944896

碩士 === 中山醫學大學 === 醫學分子毒理學研究所 === 93 === The genome of hepatitis delta virus (HDV) is composed of a circular single-stranded RNA molecule of 1.7-kb that is the smallest, and the only known circular RNA genome of the aminals RNA virus. The HDV consists of two related proteins. The small form is a 24 KDa (195 amino acids) of S-HDAg, which is essential for replication of HDV RNA genome. The large form is L-HDAg, which is a 27 KDa (214 amino acids), essential for particle assembly and inhibition of HDV RNA replication. The aim of this project is to identify cellular genes that are related to HDV replication by HDAgs induction. Therefore, we used FISH PCR technology to screen differentially expressed genes among Huh7 cells that stably expressing S-HDAg or L-HDAg. The differential expressed genes, which are induced by S-HDAg or L-HDAg were cloned by PCR and verified by autosequence. We select two genes, named C and E genes for further study. To confirm these two genes are truly HDAgs induced genes, semi-quantitative and Northern blotting are employed to quantify the expression levels of these genes. Our results indicated that the mRNA level of C gene correlated with L-HDAg expression level. Analyzed by RT-PCR, the mRNA level of E gene and is induced only by S-HDAg. Overall, we identified that both C and E genes are induced by HDAg. How the genes interact with HDAg and what role these two genes play in HDV replication need to be proved in the future.