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Infective viruses produced from full-length complementary DNA of swine vesicular disease viruses HK/70 strain

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Chinese Science Bulletin 2006年第17期51卷 2072-2078页

作者： ZHENG Haixue LIU Xiangtao SHANG Youiun WU Jinyan BAI Xingwen JIN Ye SUN Shiqi GUO Huichen TIAN Hong FENG Xia YIN Shuanghui GUO Jianhong CONG Guozheng LIU Zaixin CHANG Huiyun MA Junwu XIE Qingge Key Laboratory of Animal Virology of Ministry of Agriculture State Key Laboratory of Veterinary Etiological Biology Lanzhou Veterinary Research Institute Chinese Academy of Agricultural SciencesLanzhou 730046 China Graduate School Chinese Academy of Sciences Beijing 100081China

The full-length cDNA clone of swine vesicular disease virus HK/70 strain named pSVOK12 was constructed in order to study the antigenicity, replication, maturation and pathogenicity of swine vesicular disease virus. In vitro transcription RNA from pSVOK12 transfected IBRS-2 cells and the re- covered virus RNA were isolated and sequenced, then indirect hemagglutination test, indirect im- munofluorescence assays, eleectron microscope test, 50% tissue culture infecting dose (TCID50) assays and mouse virulence studies were performed to study the antigenicity and virulence of the recovered virus. The result showed that the infectious clones we ob- tained and the virus derived from pSVOK12 had the same biological properties as the parental strain HK/70. The full-length infectious cDNA clone, pSVOK12, will be very useful in studies of the anti- genicity, virulence, pathogenesis, maturation and replication of SVDV.

关键词：猪水泡病病毒 cDNA 传染性克隆 pSVOK12 反求遗传学

Construction and identification of reverse genetics system of Dengue type 2 virus isolated in China

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Chinese Science Bulletin 2006年第17期51卷 2065-2071页

作者： QIN Ede State Key Laboratory of Pathogen and Biosecurity Institute of Microbi- ology and Epidemiology Academy of Military Medical Sciences

To construct infectious full-length cDNA clone of dengue virus type 2 isolated in China (DEN2-43), according to the published nucleotide sequence of the virus strain, the approximately 11 kb full-length cDNAs of DEN2-43 were amplified by long RT-PCR and fusion PCR. Full-length cDNA clones were constructed by inserting the full-length cDNA into a low copy vector pWSK29, from which rescued virus D212 was acquired by transcription in vitro and electroporation. The full-length cDNA clone pD212 was infectious, and rescued virus acquired in C6/36 cells was indistinguishable from DEN2-43 virus in biological properties including suckling mice neuro- virulence. The reverse genetics system helps eluci- date the mechanism of pathogenesis of dengue virus and develop novel vaccine against dengue.

关键词：登革热病毒反求遗传学 cDNA 克隆核苷酸序列

Rescued virus from infectious cDNA clone of rabbit hemorrhagic disease virus is adapted to RK13 cells line

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Chinese Science Bulletin 2006年第14期51卷 1698-1702页

作者： LIU Guangqing NI Zheng YUN Tao ZHANG Yuying DU Qingyun SHENG Zutian LIANG Huali HUA Jionggang LI Shuangmao Chen Jianping Key Laboratory of Virology & Biotechnology of Agriculture Ministry Zhejiang Academy of Agriculture Sciences Hangzhou 310021 China Animal Veterinary Medicine College Gansu Agricultural UniversitylLanzhou 730070 China

Based on the infectious full-length cDNA clone of rabbit hemorrhagic disease virus (RHDV), the in vitro transcripts are introduced into RK13 cells. 12 h later, CPE could be observed clearly, and virual antigen could also be detected by IFA. The titre of the recovered virus is 104.6/mL. Immune electron micro-scopic observation of the virus particles revealed that the particles were rotund with a diameter of about 30 nm. Besides, virus titre quantification obtained by qRT-PCR showed a correlation between time from infection and virus titre. All these results showed that we have recovered RHDV from RK13 cells by re-verse genetics technology successfully, and this would be very useful in studies of the antigenicity, virulence, pathogenesis, maturation and new type vaccines of RHDV.

关键词：兔出血性疾病 RHDV 反求遗传学 RK13细胞 cDNA

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植物功能基因组学的研究方法及进展

烟台职业学院学报 2005年第4期11卷 93-96页

作者：牟振兴牟春红卜庆梅烟台师范学院生命科学院烟台市蓬莱市农业局

目前,基因组学的研究已从结构基因组学转向功能基因组学。植物功能基因组学的研究方法主要包括:表达序列标签(EST)、基因表达序列分析(SAGE)c、DNA微阵列和DNA芯片、反求遗传学、蛋白质组学和生物信息学;植物功能基因组学对拟南芥、水... 详细信息

目前,基因组学的研究已从结构基因组学转向功能基因组学。植物功能基因组学的研究方法主要包括:表达序列标签(EST)、基因表达序列分析(SAGE)c、DNA微阵列和DNA芯片、反求遗传学、蛋白质组学和生物信息学;植物功能基因组学对拟南芥、水稻、高粱等作物的研究取得了新的进展。

关键词：功能基因组学基因表达序列分析反求遗传学蛋白质组