| Summary: | 碩士 === 臺灣大學 === 微生物學研究所 === 95 === Human hepatitis B virus (HBV) is endemic in many places in the world, especially in Asian. It may cause serious liver diseases, like liver fibrosis, cirrhosis, and hepatocellular carcinoma (HCC). HBV cellular receptor is not yet known. Lacking of proper susceptible culture system and animal model makes it difficult to study the early process of virus infection. There are three glycoproteins in HBV surface antigens: large, middle, and small. The large protein is thought contain the cellular receptor binding site.
In my study, I use the mammalian tandem affinity purification (TAP) system to purify the recombinant large HBV surface antigen and the interaction complex, to find the recombinant large HBsAg interaction protein. There might be receptor candidates in these interaction proteins. First, purify tagged large protein expressed in mammalian cells. Then, incubate the purified recombinant proteins with human primary hepatocyte membrane proteins, which contain the cellular receptor of HBV. Use TAP system again to purify the interaction protein complexes of large protein. Utilize mass spectrometer to identify these interaction proteins.
After several experimental improvements in purification and binding assay, I find some cellular proteins interact with recombinant large protein. But it’s not found any receptor candidate in these interaction proteins. The found interaction proteins are probably related with large protein post-translational modification.
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