甜菜坏死黄脉病毒内蒙分离物RNA3序列分析及编码25kD蛋白基因在大肠杆菌中的表达

以甜菜坏死黄脉病毒（ＢＮＹＶＶ）内蒙分离物的总ＲＮＡ为模板,通过反转录－ＰＣＲ扩增获得ＢＮＹＶＶＲＮＡ３全长ｃＤＮＡ。将其克隆到ｐＧＥＭ－７Ｚｆ（＋）上,得到重组质粒ｐＧＢＹ５６。序列分析结果表明,内蒙分离物ＲＮＡ３基因组全长为１７７５ｎｔ,其中包含３个开放阅读框架,分别编码２５ｋＤ蛋白、４．６ｋＤ蛋白和一种由５９个氨基酸组成的Ｎ蛋白。与法国Ｆ２分离物、德国Ｇ１分离物和日本Ｓ分离物相比,其核苷酸序列的同源性分别为９６．４％、９６．８％和９７．３％。将２５ｋＤ蛋白编码基因克隆到ｐＪＷ２上,构建了该基因的原核表达载体。ＳＤＳ－ＰＡＧＥ和Ｗｅｓｔｅｒｎｂｌｏｔｉｎｇ分析结果表明,２５ｋＤ蛋白基因在Ｅ．ｃｏｌｉＢＬ２１（ＤＥ３）中经温度（４２℃）诱导后,可特异地表达２５ｋＤ蛋白     

兔出血症病毒结构多肽分析

粗提病毒经Ｓｅｐｈａｒｏｓｅ４Ｂ层析后,获得纯化的兔出血症病毒（ＲＨＤＶ）。提纯病毒经过ＳＤＳ－ＰＡＧＥ经考马斯亮蓝染色显示Ａ、Ｂ、Ｃ、Ｄ、Ｅ、Ｆ和Ｇ７条多肽,凝胶扫描显示Ａ为ＲＨＤＶ主要结构多肽。用多抗和单抗作免疫转印分析,证实Ａ、Ｂ、Ｃ、Ｄ、Ｅ、和Ｇ为结构多肽,此６条结构多肽间的抗原关系十分密切。     

柯萨奇B3病毒基因在重组痘苗病毒中的表达

将编码柯萨奇Ｂ３病毒（ＣＶＢ３）衣壳蛋白ＶＰ１和ＶＰ２的基因,分别克隆到具有７．５ｋ启动子的痘苗病毒表达载体ｐＧＪＰ５上;将ＣＶＢ３衣壳蛋白全基因克隆到具有Ｔ７启动子的痘苗表达载体ｐＴＭ１上,并筛先到相应的重组痘苗病毒ＶＶＰ１、ＶＶＰ２和ＶＶＰ／４／２／３／１。ＶＶＰ１和ＶＶＰ２稳定表达产物为ＣＶＢ３衣壳蛋白ＶＰ１和ＶＰ２,而ＶＶＰ４／２／３／１为一无分泌性的多聚蛋白,且这三种表达产物均属无分泌性     

减蛋综合征病毒100K蛋白基因的克隆与序列分析

用常规方法提取减蛋综合征病毒（ＥＤＳＶ）中国分离株（ＡＡ２株）病毒ＤＮＡ,分别构建了限制性内切酶ＨｉｎｄⅢ、ＳｐｈⅠ、ＰｓｔⅠ水解片段的全基因文库,并对其中１００Ｋ蛋白基因的序列进行了分析。ＥＤＳＶ１００Ｋ蛋白基因位于减蛋综合征病毒基因组５５．７～６４．８物理图谱单位（ｍ．ｕ）,共２０９１个核苷酸（ｎｔ）,其编码产物由６９６个氨基酸（ａａ）组成,推测其分子量为７７．７ｋＤ。编码蛋白氨基酸同源性分析表明,ＥＤＳＶ１００Ｋ蛋白与人腺病毒（Ａｄ２、Ａｄ５、Ａｄ１２、Ａｄ４１）、Ⅰ群禽腺病毒（ＣＥＬＯ和ＦＡＶ１０）的同源性为３２．３～３４．４％之间,而与羊腺病毒（ＯＡＶ）的同源性达到５６．４％。     

腺病毒4型E3区核苷酸序列测定及其基因结构与功能分析

本文将Ａｄ４基因组相当于７３．３－８９．２基因图谱单位的ＤＮＡ片段进行了序列测定及基因结构分析,它包括了Ａｄ４Ｅ３区全基因及该区两侧的部分序列。序列分析表明,Ａｄ４ Ｅ３区从ＴＡＴＡＡ ｂｏｘ起至该区基因结束共４７７８ｂｐ,编码１１个大于６ｋＤ的开放读码框架。对Ａｄ４ Ｅ３区ＯＲＦ分析结果表明,Ａｄ４ Ｅ３区编码的１９．３ｋ,１５ｋ,１０．４ｋ蛋白,分别与Ａｄ２ Ｅ３区的ｇｐ１９ｋ,１４．ｋ和１０     

草鱼出血病病毒多肽的免疫原性

采用中和试验比较了草鱼出血病病毒ＧＣＨＶ８７３的１１个多肽的免疫原性,确定了主要中和抗原。纯化的单个病毒多肽免疫家兔获得抗血清,多太ＶＰ１、ＶＰ５、ＶＰ６和ＶＰ９的抗血清具有中和效价,而ＶＰ２、ＶＰ３、ＶＰ４、ＶＰ８、ＶＰ１０和ＶＰ１１则不能诱生中和抗体。其中ＶＰ５的抗血清中和效价最高,因此,ＶＰ６极可能是病毒多肽中的主要中和抗原。     

球形幽门螺杆菌分子生物学研究

为研究幽门螺杆菌（ＨＰ）球形变异本质,作者通过延期培养和采用亚抑菌浓度抗生素,使３株ＨＰ发生球形变异,对弯曲形和球形ＨＰ作了ＳＤＳ－ＰＡＧＥ、免疫印迹及４个毒力基因片段ＰＣＲ和ＰＣＲ－ＳＳＣＰ分析。ＳＤＳ－ＰＡＧＥ图谱显示球形ＨＰ分子量在７４×１０４以上的蛋白含量减少,免疫印迹显示球形ＨＰ１２５×１０４蛋白条带反应减弱,而抗生素诱变的球形ＨＰ分子量为１１×１０４和６３×１０４的蛋白条带反应增强。ＰＣＲ及ＰＣＲ－ＳＳＣＰ结果表明球形ＨＰ的ｈｐａＡ,ＶａｃＡ,ＣａｇＡ和ＵｒｅＡ４个毒力基因片段未发生缺失,但在ｈｐａＡ或ＶａｃＡ基因中存在点突变     

用对流电泳提纯兔出血症病毒

兔出血症病毒（ＲＨＤＶ）在ｐＨ８．６的琼脂糖凝胶上电泳,在负极侧有一血凝峰,免疫电镜观察有大硅ＲＨＤＶ颗粒,证实ＲＨＤＶ病毒粒子带正电荷。利用这一特性可用对流电泳提纯病毒。将抗体与病毒颗粒形成的沉淀带切下,作ＳＤＳ－ＰＡＧＥ,经免疫转印出现６条区带,其中６０ｋＤ的ＶＰ_１为主要结构多肽。用免疫复合物提取核酸,以狄高辛标记制成探针,能与病毒核酸和克隆的ＲＨＤＶｃＤＮＡ２．２ｋｂ片段和４．８ｋｂ片段杂交,探针灵敏度达ｐｇ水平。能用病毒核酸作模板制备探针,证实ＲＨＤＶ的核酸为ＤＮＡ。     

蜀柏毒蛾核型多角体病毒结构多肽及基因组酶切分析

对蜀伯毒蛾核型多角体病毒（Ｐａｒｏｃｎｅｒｉａ ｏｒｉｅｎｔａ Ｎｕｃｌｅａｒ ｐｏｌｙｈｅｄｒｏｖｉｒｕｓ,简称ＰａｏｒＮＰＶ）形态结构、结构多肽、限制内切酶图谱等特性进行了研究。采用不连续系统垂直板ＳＤＳ－ＰＡＧＥ分析了ＰａｏｒＮＰＶ的多角体蛋白、病毒粒子结构多肽。应用５种限制性内切酶对ＰａｏｒＮＰＶ基因组ＤＮＡ进行了酶切分析。结果表明：经热处理的多角体蛋白仅有一条带,分子量为３１．５ｋＤ,不     

不同品种花生种子蛋白质的电泳分析

对中国花生（ＡｒａｃｈｉｓｈｙｐｏｇａｅａＬ．）５种不同类型的４６个栽培品种的种子蛋白质进行电泳分析。ＳＤＳＰＡＧＥ和ＩＥＦＳＤＳＰＡＧＥ（双向电泳）的结果显示,不同品种的花生种子蛋白多肽组成存在４种类型。与初步纯化的花生球蛋白及伴花生球蛋白的ＳＤＳＰＡＧＥ结果比较,花生种子蛋白组成的主要差异在于花生球蛋白亚基组成的不同,其中类型Ⅰ花生球蛋白主要含有４１ｋＤ、３８５ｋＤ和２个１８ｋＤ亚基,类型Ⅱ主要含４１ｋＤ、３８５ｋＤ、３７５ｋＤ和３个１８ｋＤ亚基,类型Ⅲ主要含４１ｋＤ、３８．５ｋＤ、３６．５ｋＤ和３个１８ｋＤ亚基,类型Ⅳ主要含４１ｋＤ、３８．５ｋＤ、３７５ｋＤ、３６．５ｋＤ和３个１８ｋＤ亚基。不同品种的伴花生球蛋白多肽组成基本一致。对不同类型８个品种种子蛋白的氨基酸组成进行测定,发现类型Ⅰ的含硫氨基酸含量较高。     

Entwicklungsgeschichte und Ultrastruktur von Pollenkitt und Exine bei nahe verwandten entomophilen und anemophilen Angiospermensippen derAlismataceae,Liliaceae, Juncaceae,Cyperaceae, Poaceae undAraceae

Some closely related members of the monocotyledonous familiesAlismataceae, Liliaceae, Juncaceae, Cyperaceae, Poaceae andAraceae with variable modes of pollination (insect- and wind-pollination) were studied in relation to the ultrastructure of pollenkitt and exine (amount, consistency and distribution of pollenkitt on the surface of pollen grains). The character syndromes of pollen cementing in entomophilous, anemophilous and intermediate (ambophilous or amphiphilous) monocotyledons are the same in principal as in dicotyledons. Comparing present with former results one can summarize: 1) The pollenkitt is always produced in the same manner by the anther tapetum in all angiosperm sub-classes. 2) The variable stickiness of entomophilous and anemophilous pollen always depends on the particular distribution and consistency of the pollenkitt, but not its amount on the pollen surface. 3) The mostly dry and powdery pollen of anemophilous plants always contains a variable amount of inactive pollenkitt in its exine cavities. 4) A step-by step change of the pollen cementing syndrome can be observed from entomophily towards anemophily. 5) From the omnipresence of pollenkitt in all wind-pollinated angiosperms studied one can conclude that the ancestors of anemophilous angiosperms probably have been zoophilous (i.e. entomophilous) throughout.

     

Identification and Characterization of the First Equine Parainfluenza Virus 5

正Dear Editor,Parainfluenza virus 5 (PIV5), known as canine parainfluenza virus in the veterinary field, is a negative-sense,nonsegmented, single-stranded RNA virus belonging to the Paramyxoviridae family (Chen 2018). The virus was first reported in primary monkey kidney cells in 1954 (Hsiung1972), then it has been frequently discovered in various     

Pathogenic Characterization and Full Length Genome Sequence of a Reassortant Infectious Bursal Disease Virus Newly Isolated in Pakistan

<正>Dear Editor,Infectious bursal disease (IBD) is one of the most important diseases of the poultry. The IBD virus (IBDV), a nonenveloped virus belonging to the Birnaviridae family with a genome consisting of two segments of double-stranded RNA (segments A and B), targets B lymphocytes of bursa of Fabricious leading to immunosuppression. In Pakistan,poultry farming is the second biggest industry and IBD is the second biggest disease threating the poultry sector.However, there is limited genome information of IBDV     

Similar aerosol emission rates and viral loads in upper respiratory tracts for COVID-19 patients with Delta and Omicron variant infection

Highlights
1 Aerosol emission rates of Delta or Omicron patients were similar.
2 Viral loads in upper respiratory tract of Alpha, Delta and Omicron patients were similar.
3 Viral loads in upper respiratory tract of vaccinated or unvaccinated Delta patients had no difference.     

Recombinant human interferon-α1b inhibits SARS-CoV-2 better than interferon-α2b in vitro

Highlights
1) A comprehensive evaluation method for anti-SARS-CoV-2 drugs was established based on RT-qPCR, TCID₅₀ method, and immunofluorescence.
2) A significant antiviral effect of rHuIFN-α1b was shown with EC₅₀=0.12 IU/mL in Vero cells and EC₅₀=0.52 IU/mL in Calu-3 cells, which was better than rHuIFN-α2b (EC₅₀=0.25 IU/mL in Vero cells and EC₅₀=2.48 IU/mL in Calu-3 cells).
3) rHuIFN-α1b has a good potential in the application of anti-COVID-19 therapy.