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    胡仲祺老師實驗室

    Hu, Chung-Chi
                                                
    胡仲祺 教授                                     

    Email: cchu@dragon.nchu.edu.tw

    地址:中興大學食品暨生物科技大樓6F615室

    Office phone:22840328 ext.621

    Lab phone:22840328 ext.628

    主要教授課程 (含大學部及研究所) 

    • 生物化學
    • 高等生物化學
    • 分子生物學
    • 分子生物學技術與原理
    • 生物資訊學導論
    • 生物資訊學特論

     

    學經歷

    • 學士：國立中興大學 植物病理系 1985
    • 碩士：國立中興大學 植物病理研究所 1987
    • 博士：Dept. Plant Pathology, University of Kentucky 1997
    • 博士後研究：Dept. Plant Pathology, University of Kentucky 1998
    • 助理研究員：國立中興大學 遺傳工程中心
    • 助理教授：國立中興大學 生物科技學研究所
    • 副教授：國立中興大學 生物科技學研究所 (2009.08)
    • 教授：國立中興大學 生物科技學研究所 (2016.02)

     

    研究項目簡介

    • 胡瓜嵌紋病毒衛星核酸複製及包被機制之探討與衛星核酸載體之研發：

             衛星核酸為病毒之分子寄生物，需要依賴輔助病毒進行其複製、包被、及傳播，但卻與其輔助病毒無核酸序列同源性。許多衛星核酸不帶有任何顯著的轉譯區，然而卻具有調控其輔助病毒之複製及病徵表現的能力，因此其生物活性之表達必須藉由其核酸分子本身與其他相關因子之直接交互作用，且所需之複製、包被等調控訊息亦須存在於此核酸分子上。故衛星核酸為一極佳之材料用以探討其輔助病毒之複製、包被、及病徵表現之機制。且因衛星核酸之分子遠較其輔助病毒為小，亦可在寄主植物上累積至高量，故衛星核酸具有極高的潛力可被發展成為高量表達之基因載體系統，在病毒分子生物學之基礎研究及實際應用上均具有極高的價值。

             胡瓜嵌紋病毒為cucumovirusgroup之模式病毒，寄主範圍極廣，包含單、雙子葉植物近一千種，具有極高之經濟重要性。本實驗室自中興大學徐堯煇教授處獲得十九個胡瓜嵌紋病毒株系，之中十五個株系帶有衛星核酸。而其中C株系(CMV-C)之衛星核酸(C-Sat)可減輕其輔助病毒之病徵表現，但M株系(CMV-M)之衛星核酸(M-Sat)則無影響病徵表現之能力。此二衛星核酸皆已被選殖及定序，並可經由T7起動子轉錄出具有感染力之RNA。由於此衛星核酸長度僅有334個核苷酸，因此可作為一理想之實驗系統以探討病毒或衛星核酸複製、包被所需之基本訊息，而所獲得之結果亦可作為將衛星核酸發展為基因載體系統之依據，且因胡瓜嵌紋病毒之寄主範圍極廣，預期此載體系統將可被廣泛應用於多種經濟作物上。

     

    • 台灣雙生病毒病害之診測：

             銀葉粉蝨傳播之雙生病毒 (geminiviruses)可危害40種以上蔬菜和纖維作物,發生嚴重時可完全摧毀作物的生產，此病害的發生由熱帶地區逐漸擴展至亞熱帶溫帶,已成全球性的重要植物性病毒病原。在本省已有數種作物及雜草發生類似雙生病毒感染，造成黃化及葉片捲曲的病徵，但這些病害皆有待進一步確定其病原。近期中，南部有些地區蕃茄捲葉病的肆虐，使得蕃茄幾無收成。而其媒介昆蟲(粉蝨類)亦已逐漸適應本省的農作物生態，密度急劇升高。可預見雙生病毒即將躍居台灣農作物之主要病原。利用傳統病毒學的方法鑑定雙生病毒已被證明相當困難，因此本實驗室擬建立分子生物學方法以鑑定本省所發生之雙生病毒病害，並發展分子或血清試劑，提供未來抗病育種或種苗檢疫的基本工具。

     

    • 簡化雙生病毒感染性選殖株之構築技術與新型雙生病毒潛隱性啟動子的研究：

             本團隊已研發利用滾環式擴增法構築雙生病毒感染性選殖株之簡便方法(Wu et al., 2008)，並以霍香薊黃脈病毒(Ageratum yellow vein virus, AYVV)作為研究材料，發現新型原核性啟動子，位於AYVV基因體中核苷酸位置nt 762-831的區段，並命名為AV3啟動子，其下游並有一個可以在原核生物中高量表現的短open reading frame。此新型原核啟動子在其他雙生病毒(如番茄捲葉病毒 Tomato leaf curl virus，TLCV)中也存在並有活性。此AV3啟動子具有高度活性，可在大腸桿菌中以構成性(constitutive)的方式大量表現報導基因，不需外加誘導物 (Wang et al., 2013)。後續並定位出AV3啟動子在原核與真核系統中的轉錄與轉譯起始位置(Wang et al., 2014)。此研究對於雙生病毒的演化來源與在不同寄主系統中的基因調控機制提供進一步的解釋。而此啟動子可應用於以原核或植物系統大量生產具有高經濟價值的外源蛋白(如疫苗或醫用蛋白)或作為選殖載體等方面。此成果與研究啟動子的經驗亦可用於進行寄主因子之選殖、表現、以及miRNA啟動子的分析。 
       

    期刊論文

    1. Kuo, S. Y., Lin, Y. C., Lai, Y. C., Liao, J. T., Hsu, Y. H., Huang, H. C., and Hu, C. C. (2018) Production of fluorescent antibody-labeling proteins in plants using a viral vector and the application in the detection of Acidovorax citrulli and Bamboo mosaic virus. PloS ONE, 13, e0192455.
    2. Huang, Y. W., Hu, C. C., Tsai, C. H., Lin, N. S., and Hsu, Y. H. (2017) Chloroplast Hsp70 isoform is required for age-dependent tissue preference of Bamboo mosaic virus in mature Nicotiana benthamiana leaves. Molecular Plant-Microbe Interactions, 30, 631-645.
    3. Chang, K. C., Chang, L. T., Huang, Y. W., Lai, Y. C., Lee, C. W., Liao, J. T., Lin, N. S., Hsu, Y. H., and Hu, C. C. (2017) Transmission of Bamboo mosaic virus in bamboos mediated by insects in the order Diptera. Frontiers in Microbiology, 8, 870.
    4. Chen, T. H., Hu, C. C., Liao, J. T., Lee, Y. L., Huang, Y. W., Lin, N. S., Lin, Y. L., and Hsu, Y. H. (2017) Production of Japanese encephalitis virus antigens in plants using Bamboo mosaic virus-based vector. Frontiers in Microbiology, 8, 788.
    5. Liou, M. R., Hu, C. C., Chou, Y. L., Chang, B. Y., Lin, N. S., and Hsu, Y. H. (2015) Viral elements and host cellular proteins in intercellular movement of Bamboo mosaic virus. Current Opinion in Virology, 12, 99-108.
    6. Muthamilselvan, T., Lee, C. W., Cho, Y. H., Wu, F. C., Hu, C. C., Liang, Y. C., Lin, N. S., and Hsu, Y. H. (2015) A transgenic plant cell-suspension system for expression of epitopes on chimeric Bamboo mosaic virus particles. Plant Biotechnology Journal, 14, 231-239.
    7. Hung, C. J., Huang, Y. W., Liou, M. R., Lee, Y. C., Lin, N. S., Meng, M., Tsai, C. H., Hu, C. C., and Hsu, Y. H. (2014) Phosphorylation of coat protein by protein kinase CK2 regulates cell-to-cell movement of Bamboo mosaic virus through modulating RNA binding. Molecular Plant-Microbe Interactions, 27, 1211-1225.
    8. Wu, C. C., Huang, W. C., and Hu, C. C. (2014) An ultrasensitive label-free electrochemical impedimetric DNA biosensing chip integrated with a DC-biased AC electroosmotic vortex. Sensors and Actuators B: Chemical, 209, 61-68.
    9. Wang, I. N., Hu, C. C., Lee, C. W., Yen, S. M., Yeh, W. B., Hsu, Y. H., and Lin, N. S. (2014) Genetic diversity and evolution of satellite RNAs associated with the Bamboo mosaic virus. PLoS One, 9, e108015.
    10. Wang, W. C., Wu, C. Y., Lai, Y. C., Lin, N. S., Hsu, Y. H., and Hu, C. C. (2014) Characterization of the cryptic AV3 promoter of ageratum yellow vein virus in prokaryotic and eukaryotic systems. PLoS One, 9, e108608.
    11. Hung, C. J., Hu, C. C., Lin, N. S., Lee, Y. C., Meng, M., Tsai, C. H., and Hsu, Y. H. (2013) Two key arginine residues in the coat protein of Bamboo mosaic virus differentially affect the accumulation of viral genomic and subgenomic RNAs. Molecular Plant Pathology, 15, 196-210.
    12. Liou, M. R., Huang, Y. W., Hu, C. C., Lin, N. S., and Hsu, Y. H. (2013) A dual gene-silencing vector system for monocot and dicot plants. Plant Biotechnology Journal, 12, 330-343.
    13. Wang, W. C., Hsu, Y. H., Lin, N. S., Wu, C. Y., Lai, Y. C., and Hu, C. C. (2013) A novel prokaryotic promoter identified in the genome of some monopartite begomoviruses. PLoS One, 8, e70037.
    14. Huang, Y. W., Hu, C. C., Lin, N. S., and Hsu, Y. H. (2012) Unusual roles of host metabolic enzymes and housekeeping proteins in plant virus replication. Current Opinion in Virology, 2, 676-682.
    15. Chen, T. H., Chen, T. H., Hu, C. C., Liao, J. T., Lee, C. W., Liao, J. W., Lin, M. Y., Liu, H. J., Wang, M. Y., Lin, N. S., and Hsu, Y. H. (2012) Induction of protective immunity in chickens immunized with plant-made chimeric Bamboo mosaic virus particles expressing very virulent Infectious bursal disease virus antigen. Virus Research, 166, 109-115.
    16. Huang, Y. W., Hu, C. C., Liou, M. R., Chang, B. Y., Tsai, C. H., Meng, M., Lin, N. S., and Hsu, Y. H. (2012) Hsp90 interacts specifically with viral RNA and differentially regulates replication initiation of Bamboo mosaic virus and associated satellite RNA. PLoS Pathogens, 8, e1002726.
    17. Prasanth, K. R., Huang, Y. W., Liou, M. R., Wang, Y. L., Hu, C. C., Tsai, C. H., Meng, M., Lin, N. S., and Hsu, Y. H. (2011) Glyceraldehyde 3-phosphate dehydrogenase negatively regulates the replication of Bamboo mosaic virus and its associated satellite RNA. Journal of Virology, 85, 8829-8840.
    18. Cheng, S. F., Huang, Y. P., Wu, Z. R., Hu, C. C., Hsu, Y. H., and Tsai, C. H. (2010) Identification of differentially expressed genes induced by Bamboo mosaic virus infection in Nicotiana benthamiana by cDNA-amplified fragment length polymorphism. BMC Plant Biology, 10:286.
    19. Huang, Y. W., Hu, C. C., Lin, N. S., and Hsu, Y. H. (2010) Mimicry of molecular pretenders: the terminal structures of satellites associated with plant RNA viruses. RNA Biology, 7, 162-171.
    20. Hu, C. C., Hsu, Y. H., and Lin, N. S. (2009) Satellite RNAs and satellite viruses of plants. Viruses. 1, 1325-1350.
    21. Tseng, Y. H., Hsu, H. T., Chou, Y. L., Hu, C. C., Lin, N. S., Hsu, Y. H., and Chang, B. Y. (2009) The two conserved cysteine residues of the triple gene block protein 2 are critical for both cell-to-cell and systemic movement of Bamboo mosaic virus. Molecular Plant-Microbe Interactions, 22, 1379-1388.
    22. Liao, J. Y., Hu, C. C., Chen, C. C., Chang, C. H., and Deng, T. C. (2009) Full-length sequence analysis of a distinct isolate of Bidens mottle virus infecting sunflower in Taiwan. Arch. Virol. 154, 723-725.
    23. Huang, Y. W., Hu, C. C., Lin, C. A., Liu, Y. P., Tsai, C. H., Lin, N, S., and Hsu, Y. H. (2009) Structural and functional analyses of the 3′ untranslated region of Bamboo mosaic virus satellite RNA. Virology. 386, 139-153.
    24. Huang, Y. W., Hu, C. C., Lin, N. S., Tsai, C. H., and Hsu, Y. H. (2008) In vitro replication of Bamboo mosaic virus satellite RNA. Virus Res. 136, 98-106.
    25. Shien , J. H., Wang, Y. S., Chen, C. H., Shieh, H. K., Hu, C. C., and Chang, P. C. (2008) Identification of sequence changes in live attenuated goose parvovirus vaccine strains developed in Asia and Europe. Avian Pathol. 37,499-505.
    26. Wu, C. Y., Lai, Y. C., Lin, N. S., Hsu, Y. H., Tsai, H. T., Liao, J. Y., and Hu, C.C. (2008) A simplified method of constructing infectious clones of begomovirus employing limited restriction enzyme digestion of products of rolling circle amplification. J. Virol. Methods 147, 355-359.
    27. Wu, C. Y., Yang, S. H., Lai, Y. C., Lin, N. S., Hsu, Y. H., and Hu, C. C. (2007) Unit-length, single-stranded circular DNAs of both polarity of begomoviruses are generated in Escherichia coli harboring phage M13-cloned begomovirus genome with single copy of replication origin. Virus Res. 125, pp. 14-28.
    28. Wu, J. R., Shien, J. H., Shieh, K. H., Hu, C. C., Gong, S. R., Chen, L. Y., and Chang, P. C. (2007) Cloning of the gene and characterization of the enzymatic properties of themonomeric alkaline phosphatase (PhoX) from Pasteurella multocida strainX-73. FEMS Microbiol. Lett. 267, 113-120.
    29. Liao, J. Y., Hu, C. C., Kao, J. L., and Deng, T. C. (2007) Identification of Tobacco mosaic virus infecting Rehmannia glutinosa. Plant Pathol. Bull. 16, 61-69.
    30. Liao, J. Y., Hu, C. C., Lin, T. K., Chang, C. A., and Deng, T. C. (2007) Identification of Squash leaf curl Philippines virus on Benincasa hispida in Taiwan. Plant Pathol. Bull. 16, 11-18.
    31. Hajimorad, M. R., Hu, C. C., and Ghabrial, S. A. (1999) Molecular characterization of an atypical Old World strain of peanut stunt virus. Arch. Virol. 144: 1587-1600.
    32. Chen, C. C., and Hu, C. C. (1999) Purification and characterization of a cucumovirus from Lisianthus rusellianus. Plant Prot. Bull. 41: 179-198.
    33. Di, R., Hu, C. C., and Ghabrial, S. A. (1999) Complete nucleotide sequence of bean pod mottle comovirus RNA1: sequence comparisons and evolutionary relationships to other comoviruses. Virus Genes 18, 129-137.
    34. Hu, C. C., and Ghabrial, S. A. (1998). Molecular evidence that strain BV-15 of peanut stunt cucumovirus is a reassortant between subgroup I and II strains. Phytopathology 88: 92-97.
    35. Hu, C. C., Sanger, M., and Ghabrial, S. A. (1998) Production of infectious RNA transcripts from full-length cDNA clones representing two subgroups of peanut stunt virus strains: mapping satellite RNA support to RNA1. J. Gen. Virol. 79, 2013-2021.
    36. Hu, C. C., Sanger, M. P., and Ghabrial, S. A. (1999) A novel defective RNA associated with peanut stunt virus strain B. (manuscript in preparation)
    37. Hu, C. C., Aboul-Ata, A. E., Naidu, R. A., and Ghabrial, S. A. (1997) Evidence for the occurrence of two distinct subgroups of peanut stunt cucumovirus strains: molecular characterization of RNA3. J. Gen. Virol. 78: 929-939.
    38. Naidu, R. A., Hu, C. C., Pennington, R. E., and Ghabrial, S. A. (1995) Differentiation of eastern and western strains of peanut sutnt cucumovirus based on satellite RNA support and nucleotide sequence homology. Phytopathology 85: 502-507.
    39. Hu, C. C., and Ghabrial, S. A. (1995) The conserved, hydrophilic and arginine-rich N-terminal domain of cucumovirus coat proteins contributes to their anomalous electrophoretic mobilities in sodium dodecylsulfate-polyacrylamide gels. J. Virol. Methods 55: 367-379.
    40. Hsu, Y. H., Wu, C. W., Lee, C. W., Hu, C. C., and Lin, F. Z. (1994) Satellite defense as a control strategy for cucumber mosaic virus. Plant Pathol. Bull. 3: 72-77.820418B005091B03.
    41. Lin, N. S., Lin, B. Y., Lo, N. W., Hu, C. C., Chow, T. Y., and Hsu, Y. H. (1994) Nucleotide sequence of the genomic RNA of bamboo mosaic potexvirus. J. Gen. Virol. 75: 2513-2518.
    42. Hsu, Y. H., Lin, F. Z., Hu, C. C., and Yin, S. C. (1989) Host reaction, serology and RNA pattern of cucumber mosaic virus isolates. Plant Brot. Bull. 31:51-59.
    43. Hsu, Y. H., Hu, C. C., Lin, N. S., and Chiu, R. J. (1988) Symptom determinant of two Taiwan strains of cucumber nmosaic virus is on RNA3. Bot. Bull. Acad. Sinica. 29: 231-237.760201b0051.