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1.
香榧绿藻Chlorella sp.是一种附着在香榧Torreya grandis ‘Merrillii’枝、叶上的藻类。为探明香榧绿藻的种属类别及亲缘关系,探究香榧绿藻的生物学特性,以3个不同地区的香榧树上采集的香榧绿藻为试验对象,通过室内培养,观察香榧绿藻的形态特征及生活习性,提取香榧绿藻DNA并进行18S rDNA序列和内转录间隔区（ITS）基因序列特异性扩增,最终序列进行BLAST比对,绘制系统进化树。结果表明:香榧绿藻由A组和B组2种不同形态的绿藻组成,不同地区香榧绿藻形态存在差异,但主要以A组形态绿藻为主。根据形态学鉴定结果表明:A组香榧绿藻与引起番茄绿藻病的椭圆小球藻Chlorella ellipsoidea形态相近,B组香榧绿藻与引起黄瓜绿藻病的丝藻Ulothrix sp.形态相近;但香榧绿藻对pH值的反应规律与斜生栅藻Scenedesmus obliquus相近。通过序列比对发现,2种形态的香榧绿藻的18S rDNA和ITS区基因序列也与栅藻科Scenedesmace的Asterarcys quadricellulare的同源性高,相似率达100%。结合传统形态学方法和分子生物学技术,将A组香榧绿藻初步鉴定为栅藻科的Asterarcys quadricellulare,B组香榧绿藻为其变种,且2组香榧绿藻与同属于栅藻科的Scenedesmus sp. SM15_4（KT778094.1）的ITS区基因序列相同。香榧绿藻具有较好的耐碱性,生长周期长且不容易腐烂。这些生物学特性赐予香榧绿藻在香榧树上长期寄生的能力。 相似文献
2.
鸡传染性贫血病PCR试剂盒技术的研究及应用 总被引：2，自引：0，他引：2
根据鸡传染性贫血病毒（CIAV）的VP1、VP2基因序列的结构特点,设计合成了一对引物,建立了检测鉴别鸡传染性贫血病毒PCR技术,并研制出CIAV—PCR检测试剂盒。试验结果显示,该CIAV—PCR检测试剂盒对参考菌株和地方分离株均能特异性地扩增出420bp条带,而对其他禽病病原体的扩增结果为阴性;该CIAV—PCR试剂盒最低能检测出10fg的CIAV DNA模板;保存期测定结果显示,在-20℃条件下保存至1、3．6、9个月时,试剂盒的敏感性无明显变化,仍能检测到10～100fg的CIAV DNA模板,保存至12个月时其敏感度虽然降低了1个滴度,但仍能100％检出人工感染鸡的临床样品。表明该CIAV—PCR检测试剂盒对CIAV临床样品的检测具有特异、敏感、快速和准确的特点,适合在临床生产中推广使用。 相似文献
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根据GenBank中已发表的鸭瘟病毒TK基因序列,设计一对引物,对1株鸭瘟病毒强毒和1株鸭瘟病毒疫苗毒进行PCR扩增。将扩增的目的片段分别克隆到pMD18-T载体,经EcoRⅠ和HindⅢ双酶切鉴定,获得阳性重组质粒,然后对阳性重组质粒进行序列测定及分析。结果表明,本实验所扩增的鸭瘟病毒TK基因及侧翼UL24基因大小为1 995 bp,鸭瘟强、弱毒株TK基因及侧翼UL24基因序列完全相同,该病毒TK基因与鸭瘟病毒其它毒株AY911509与AY963569,四川株DQ640611,AV1221株EF173464,sd-01株EF417996同源性分别为99.5%,99.9%,100%,99.9%,100%,而该病毒UL24基因与鸭瘟病毒其它毒株AY911511,DQ227739,EF417996同源性为99.9%,99.8%,99.9%,表明鸭瘟病毒强弱毒株TK基因及侧翼UL24基因高度保守。为构建鸭瘟病毒TK基因缺失的转移载体奠定了基础。 相似文献
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甘肃省河西地区菜豆病毒病分子检测 总被引：1，自引：1，他引：0
以采集自甘肃省河西地区表现病毒病症状的菜豆病株叶组织总RNA为模板,进行RT-PCR扩增,鉴定出苜蓿花叶病毒、小西葫芦黄花叶病毒、菜叶普通花叶病毒、黑眼豇豆花叶病毒4种病毒.对预期大小的扩增产物进行直接测序.结果表明:该地区菜豆病毒病的发生以苜蓿花叶病毒、黑眼豇豆花叶病毒和小西葫芦黄花叶病毒复合侵染为主. 相似文献
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配制不同浓度的猪粪浸提液,自然放置,以观察在猪粪有机污水中适宜生长的浮游藻类。结果表明,不同浓度的污水经过一定时间均有藻类生长,并且形成水华。各浓度适合生长的优势藻类不同,100%的猪粪污水主要生长衣藻(Chlamydomonassp.),50%的猪粪污水主要生长衣藻(Chlamydomonassp.)和小球藻(Chlorellasp.),30%的猪粪污水主要生长小球藻(Chlorellasp.),10%的猪粪污水主要生长纤维藻(Ankistrodesmussp.)。藻类最大生物量随浓度升高而降低,呈现高浓度抑制的趋势。水体氮磷营养盐随藻类的生长繁殖而快速下降,从而限制藻类继续增殖,藻类数量下降,水华消失。 相似文献
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以蛋白核小球藻为受试物,分别设0(CK)、0.06、0.12、0.18、0.24、0.30、0.36 mg/L十六烷基三甲基溴化铵(CTAB)处理,考察CTAB对藻生长和生化指标的影响,分析其致毒机理。结果表明:CTAB对蛋白核小球藻96 h EC50值为0.17 mg/L;藻细胞叶绿素a、水溶性蛋白质含量及超氧化物歧化酶(SOD)活性随CTAB浓度增加呈先升后降趋势,当CTAB剂量≤0.06 mg/L时,叶绿素a含量略微上升,>0.06 mg/L时,叶绿素a含量急剧下降;CTAB剂量≤0.24 mg/L处理下蛋白质含量增加,CTAB剂量≥0.30 mg/L处理时水溶性蛋白含量低于对照组;除剂量≤0.06 mg/L处理外,SOD活性随CTAB剂量的增大呈下降趋势;脂质过氧化丙二醛(MDA)含量则随CTAB浓度增加逐渐上升;CTAB对蛋白核小球藻的致毒机理为通过破坏细胞膜完整性,抑制SOD活性,导致MDA含量持续上升。 相似文献
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以间接免疫荧光抗体技术（ＩＦＡ）为基础，并加以改进，建立了间接免疫荧光染色空斑计数技术（ＰＣＴＩＩＦＳ），用其检测鸡马立克氏病（ＭＤ）二价疫苗，可快速准确地计数疫苗中不同血清型病毒各自的空斑形成单位（ＰＦＵ）及其ＰＦＵ总数。克服了常规法不能计数二价苗中两种病毒各自空斑数的缺点，敏感性显著高于常规病毒空斑计数法。 相似文献
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While it is widely appreciated that infection with a virulent virus can produce disease in an animal, the ability of a mixture of avirulent viruses to produce disease by means of complementation or recombination in vivo has not been established. In this study, two weakly neuroinvasive herpes simplex virus type 1 (HSV-1) strains were simultaneously inoculated onto the footpads of mice. Many (62%) of the animals that received a 1:1 mixture of the viruses died, whereas the animals that received a similar or 100-fold higher dose of each agent alone survived. Of fourteen viruses isolated from the brains of ten mice that died after receiving the mixture of the two weakly neuroinvasive viruses, eleven were recombinants; three of these recombinants were lethal when reapplied to the footpads of mice. These results show that two avirulent HSV-1 variants may interact in vivo to produce virulent recombinants and a lethal infection. They also suggest that different genetic lesions account for the weakly neuroinvasive character of the HSV-1 strains ANG and KOS after footpad inoculation. 相似文献
10.
[目的]选择合适的小球藻种属鉴定基因,对高产油微藻进行分子鉴定。[方法]从分子鉴定常用的4条基因[核基因组rDNA的18S rRNA gene、内转录间隔区（ITS）、内转录间隔区2（ITS-2）和叶绿体rbcL基因]中筛选出最为适用的小球藻鉴定基因,并对从自然水体中分离筛选出的5株油脂含量在30%以上的小球藻（Chlorellasp.）进行了分类鉴定。[结果]ITS序列变异程度高,片段长度短,序列长度在种间变异较大,而在小球藻种内高度保守,且种间距离（0.439 6±0.135 9）远大于种内距离（0.045 7±0.084 3）,适用于小球藻属内种的鉴定。应用ITS序列分别将5株高产油藻鉴定为1株C.vulgaris、2株C.sorokiniana藻和2株Chlorella sp.。[结论]为建立藻类的鉴定基因库提供了参考。 相似文献
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[目的]选择合适的小球藻种属鉴定基因,对高产油微藻进行分子鉴定。[方法]从分子鉴定常用的4条基因核基因组rDNA的18SrRNAgene、内转录间隔区（ITS）、内转录间隔区2（ITS2）和叶绿体rbcL基因中筛选出最为适用的小球藻鉴定基因,并对从自然水体中分离筛选出的5株油脂含量在30%以上的小球藻（Chlorellasp.）进行了分类鉴定。[结果]ITS序列变异程度高,片段长度短,序列长度在种间变异较大,而在小球藻种内高度保守,且种间距离（0.4396±0.1359）远大于种内距离（0.0457±0.0843）,适用于小球藻属内种的鉴定。应用ITS序列分别将5株高产油藻鉴定为1株C.vulgaris、2株C.sorokiniana藻和2株Chlorellasp.。[结论]为建立藻类的鉴定基因库提供了参考。 相似文献
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Viruses have been hypothesized to control blooms of Aureococcus anophagefferens gen. et sp. nov. (Chrysophyceae), a marine phytoplankton that since 1985 has caused devastating summer blooms called "brown tide." By means of ultrafiltration methods, viruses specific to this alga were isolated from both the Great South Bay and Peconic Bay systems of Long Island, New York, during the summer bloom period of 1992. Cell lysis of healthy algal cultures was demonstrated, as well as continuing reinfection with serial transfers of cultures. Electron microscope surveys yielded images of phage-like virus particles with tails that could attach to A. anophagefferens cells within minutes of exposure. The isolation and cultivation of this virus highlights the need for further study of viral infection of eukaryotic algae and the potential for a better understanding of algal bloom control by viral infection. 相似文献
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依据Gen Bank中注册的鸡马立克氏病病毒(MDV)、鸡传染性喉气管炎病毒(ILTV)、鸡痘病毒(FPV)基因组核苷酸序列中的保守序列设计引物,3种病毒各设计3对引物,共计9对引物,经生物学软件筛选后,得到3对相互匹配较佳的引物。3对匹配引物经单一PCR验证后,优化复合PCR反应条件,确定最佳的退火温度、引物浓度和Taq DNA聚合酶浓度;经特异性、敏感性试验及简化PCR试验,建立简易复合PCR检测方法。复合PCR扩增出的3条带大小与预期一致,即228 bp(MDV)、400 bp(ILTV)、499 bp(FPV);建立的复合PCR方法能同时检测出100 fg MDV、ILTV、FPV的DNA。同时依据PCR技术原理,将经典的三温热循环改进为二温热循环试验,即将退火和延伸合并为一步(62℃),缩短了反应时间。建立的3种病毒的复合PCR检测方法具有较高的特异性和敏感性,可以用于临床病料的快速诊断。 相似文献
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中国部分地区高致病性猪繁殖与呼吸综合征病毒的分子流行病学研究 总被引：12，自引：1，他引：11
【目的】对来自2006年猪病流行爆发地的组织样品进行多种病毒的检测,确定引起本次流行的致病病原及PRRSV的分子流行病学特征。【方法】利用（RT-）PCR对采集的样品进行PRRSV北美型和欧洲型、猪圆环病毒2型（PCV2）、猪血凝性脑脊髓炎病毒（PHEV）、日本乙型脑炎病毒（JEV）以及瘟病毒的检测,对PRRSV阳性的进行ORF5的序列测定并分析。【结果】检测到16份PRRSV北美型阳性样品,PCV2阳性样品2份,其它病毒均未检到。PRRSV ORF5序列分析表明笔者分离到的PRRSV株可分为2个群。【结论】（1）PRRSV与2006年肆虐中国养猪业的猪病流行有直接关系;（2）2006年PRRSV流行株主要是以JX0612株为代表,但不同遗传特征的PRRSV也在流行。 相似文献
15.
Reverse transcriptase in normal rhesus monkey placenta 总被引：5，自引：0，他引：5
16.
Mapping of herpes simplex virus-1 neurovirulence to gamma 134.5, a gene nonessential for growth in culture 总被引：49，自引：0，他引：49
The gene designated gamma 134.5 maps in the inverted repeats flanking the long unique sequence of herpes simplex virus-1 (HSV-1) DNA, and therefore it is present in two copies per genome. This gene is not essential for viral growth in cell culture. Four recombinant viruses were genetically engineered to test the function of this gene. These were (i) a virus from which both copies of the gene were deleted, (ii) a virus containing a stop codon in both copies of the gene, (iii) a virus containing after the first codon an insert encoding a 16-amino acid epitope known to react with a specific monoclonal antibody, and (iv) a virus in which the deleted sequences were restored. The viruses from which the gene was deleted or which carried stop codons were avirulent on intracerebral inoculation of mice. The virus with the gene tagged by the sequence encoding the epitope was moderately virulent, whereas the restored virus reacquired the phenotype of the parent virus. Significant amounts of virus were recovered only from brains of animals inoculated with virulent viruses. Inasmuch as the product of the gamma 134.5 gene extended the host range of the virus by enabling it to replicate and destroy brain cells, it is a viral neurovirulence factor. 相似文献
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Russell CA Fonville JM Brown AE Burke DF Smith DL James SL Herfst S van Boheemen S Linster M Schrauwen EJ Katzelnick L Mosterín A Kuiken T Maher E Neumann G Osterhaus AD Kawaoka Y Fouchier RA Smith DJ 《Science (New York, N.Y.)》2012,336(6088):1541-1547
Avian A/H5N1 influenza viruses pose a pandemic threat. As few as five amino acid substitutions, or four with reassortment, might be sufficient for mammal-to-mammal transmission through respiratory droplets. From surveillance data, we found that two of these substitutions are common in A/H5N1 viruses, and thus, some viruses might require only three additional substitutions to become transmissible via respiratory droplets between mammals. We used a mathematical model of within-host virus evolution to study factors that could increase and decrease the probability of the remaining substitutions evolving after the virus has infected a mammalian host. These factors, combined with the presence of some of these substitutions in circulating strains, make a virus evolving in nature a potentially serious threat. These results highlight critical areas in which more data are needed for assessing, and potentially averting, this threat. 相似文献
19.
为掌握近年来H9亚型禽流感的流行状况和遗传变异规律,从河南、陕西发病养殖场采集病料,用鸡胚接种法分离病毒,并用RT-PCR法对分离出的H9N2AIV的HA基因进行扩增、克隆、测序及对得到的序列进行同源性和遗传性分析。结果表明:获得了2株H9N2亚型流感病毒,这2株病毒HA基因变异程度不大,核苷酸序列和氨基酸同源性分别为97.1%和97.8%,均属于BJ/1/94分支(Y280小分支)。从分子水平分析,这2株病毒均属于低致病力的H9N2禽源流感病毒。 相似文献
20.
Natural variation of canine parvovirus 总被引：20，自引：0，他引：20
C R Parrish P H O'Connell J F Evermann L E Carmichael 《Science (New York, N.Y.)》1985,230(4729):1046-1048
Canine parvovirus was first recognized during 1978. Analysis of isolates collected since its emergence revealed that viruses circulating after 1980 were antigenically different from earlier isolates. Monoclonal antibodies clearly distinguished the two strains, some being specific for either the old or the new viruses. Restriction enzyme analysis of viral DNA's showed that the post-1980 viruses were similar to earlier isolates, but some restriction site differences were present in the new strain. These results suggest that the canine parvoviruses infecting dogs in the seven areas of the United States that were sampled derive from a variant virus that replaced the original strain during 1980. 相似文献