Molecular cloning and nucleotide sequence of classical swine fever virus strain Shimen

According to the previously published CSFV sequences, 18 paris of partially overlapping primers which span the entire genome of CSFV strain Shimen were designed and synthesized. Each cDNA fragment of strain Shimen was amplified by RT-PCR method from the anticoagulant blood of strain Shimen infected pig. The PCR fragments were cloned into pGEM-T vector respectively and sequenced. The results show that we have obtained the nucleotide sequence of strain Shimen. The viral RNA consists of 12 297 nucleotides including noncoding regions of 373 and 227 bases at the 5′ and 3′ end, respectively, and a single large open reading frame spanning 11 697 nucleotides in the middle, which encodes an amino acid sequence of 3 989 residues with a calculated molecular weight of 437.6×10³. The precisely sequencing of 5′ and 3′ termini is undertaking. Supported by the National Pandeng Project Huang Qianhua: born in 1968. Graduate student     

免疫沉淀法分离纯化感染细胞中的甲肝病毒RNA

本研究采用抗血清（多抗血清）沉淀甲肝病毒,并用盐酸胍酸性酚,氯仿一步法分离纯化病毒ＲＮＡ．此方法能排除非病毒ＲＮＡ的污染,操作简单,要求条件较低,ＲＮＡ的纯度和完整性都较好     

有限稀释法筛选重组AcNPV病毒

有限稀释法筛选重组ＡｃＮＰＶ病毒钟劲胡美浩１）（北京大学生命科学学院,北京,１００８７１）关键词重组病毒;梯度稀释;筛选;测活中图分类号Ｑ３３２０引言昆虫杆状病毒表达系统是一高效表达外源基因的表达系统。用于基因工程表达外源基因的ＡｃＮＰＶ（ａｕｔｏｇ...     

痰和咽拭子PCR及血清学联合检测肺炎衣原体急性呼吸道感染

了解成人呼吸道感染患者急性肺炎衣原体感染的发病情况。呼吸道感染患者１１０例,同时采集痰和咽拭子标本,应用聚合酶链式反应（ＰＣＲ）检测肺炎衣原体ＤＮＡ及采静脉血检测肺炎衣原体抗体。本组患者１７例（１６％）有肺炎衣原体近期感染的急性抗体,１２例（１１％）痰和（或）咽拭子肺炎衣原体ＰＣＲ检测阳性,联合应用两种方法的阳性率为２３％（２５／１１０例）。肺炎衣原体急性感染以哮喘急性发作、肺炎、ＣＯＰＤ急性加重和急性支气管炎患者多见（分别为５７％、３５％、２６％和２５％）,其临床表现无特征性。本组成人呼吸道感染患者肺炎衣原体急性感染的发病率较高,提示肺炎衣原体是呼吸道感染的重要致病原。     

植物病毒增殖的超循环理论与抗病毒植物基因工程

考虑了两个典型的植物病毒ＴＭＶ和ＰＶＹ,从病毒基因组的复制,翻译和组装各个环节的反应方程出发得到了病毒增殖的动力学方程。这组方程是Ｅｉｇｉｎ超循环理论在病毒ＲＮＡ增殖系统中的应用和发展。     

黑胸大蠊浓核病毒的病理学研究

应用组织化学方法以及电镜超薄切片技术对黑胸大蠊浓核病毒感染蜚蠊的病理学作了初步研究．经孚尔根—亮绿染色、甲基绿—派洛宁染色、吖啶橙染色,发现蜚蠊中肠柱状上皮、盲囊、马氏管和砂囊组织不被蜚蠊浓核病毒感染外,其它几乎所有蜚蠊组织都被病毒感染,并在细胞核内形成孚尔根反应强阳性,派洛宁和吖啶橙好染的均匀物质团．电镜下发现,感染细胞核明显膨大,核内异染色质浓缩并被推向核的边缘,核仁分离并最后消失．核内病毒粒子从发生基质形成、释放,最后使核膜破裂而进入细胞质中．在少量细胞中可见到大片晶格排列的病毒颗粒．感染细胞内线粒体、内质网均退化,空泡化     

内联网络安全的研究与探讨

本文论述了当前内联网络网络安全工作对于国家安全和经济建设的重要意义、它所面临的种种威胁、网络安全体系功能需求，并阐述网络安全技术最新发展，给出了一个较为完备的安全体系可以采用的各种加强手段，包括防火墙、加密认证、网络安全扫描、入侵检测等技术。     

牛泡沫病毒BFV3026 gag基因的克隆及其在大肠杆菌中的表达

从牛泡沫病毒BFV3026感染的胎牛肺细胞中提取Hirt DNA,PCR扩增BFV3026　gag基因。序列分析确证后，将其克隆于原核表达载体pET32A，重组质粒转化大肠杆菌BL21（DE3），经IPTG诱导，使Gag蛋白得以高效表达。SDS－PAGE及Western blot证实：表达蛋白占菌体总蛋白的40%，本工作的完成为Gag蛋白功能的研究奠定基础。     

Identification of antiviral- relevant genes in the cultured fish cells induced by UV-inactivated virus

~~Identification of antiviral- relevant genes in the cultured fish cells induced by UV-inactivated virus~~     

N-terminal of L protein of vesicular stomatitis virus contains a new signal sequence

The L protein (241kD) of vesicular stomatitis virus (VSV) is the most important snbnnit of the replication complex. The existence of specific localization signal in the L protein was investigated by making recombinant constructs expressing truncated mutants of the L protein fused to green fluorescent protein (GFP) in transient transfection assays. The chimeric genes encoding varied N-terminal of L and GFP gene were put under the control of T7 promoter or CMV promoter. The fusion proteins were transiently expressed in BHK-21, COS-7, CHO or Hep G2 cells. When more than 120 residues were deleted or only 96 residues were kept on the N-terminal, the fusion proteins were shown to be distributed throughout the cells, cytoplasm and nucleus under the confocal microscope. However, other chimeric proteins with 120 or more amino acids were dotted and distributed in the perinuclear regions. And the fusion protein with 96—120 aa has the similar distribution. A thirteen-residue peptide QGYSFLHEVDKEA (108—120) was identified as localization signal, whose function would be absolutely distributed with the deficiency of D or V. Our results show that there is an independent localizing signal in N-terminal domain of L protein of VSV and this functional signal is conserved in different cell lines.