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通过RNA杂交确定的人类轮状病毒之间的遗传相关性。

Genetic relatedness among human rotaviruses as determined by RNA hybridization.

作者信息

Flores J, Perez I, White L, Perez M, Kalica A R, Marquina R, Wyatt R G, Kapikian A Z, Chanock R M

出版信息

Infect Immun. 1982 Aug;37(2):648-55. doi: 10.1128/iai.37.2.648-655.1982.

Abstract

Viral RNAs from human rotaviruses were compared by gel electrophoresis and by hybridization to probes prepared by in vitro transcription of two well-characterized laboratory strains (Wa and DS-1). Also, the viral RNAs were compared by hybridization to probes prepared from three of the test viruses. Thirteen specimens (diarrheal stools) were obtained from infants and children 5 to 21 months old on a single day at the emergency ward of the Caracas Children's Hospital, and an additional specimen was obtained from the same hospital 6 months before. When the electrophoresed viral RNAs were stained with ethidium bromide and examined by UV light, five different migration patterns (electropherotypes) were distinguished on the basis of differences in mobility of the RNA segments. The hybridization technique that was employed permitted only qualitative comparisons of corresponding genes of different human rotaviruses. Ten of the specimens contained enough virus to yield sufficient RNA for hybridization studies. Eight of the viruses studied by hybridization contained 4 to 11 genes that reacted specifically with the Wa probe to yield double-stranded RNA segments with a mobility similar to that of Wa viral RNA or test virus RNA. The other two viruses contained 11 genes that reacted specifically with the DS-1 hybridization probe to yield double-stranded RNA segments with a mobility similar to DS-1 viral RNA or test virus RNA. A more complex picture emerged when hybridization probes were prepared from three of the test viruses and used to compare the different electropherotypes. Corresponding genes that exhibited similar migration did not necessarily exhibit homology when studied by hybridization. Also, some corresponding genes that exhibited homology did not have the same mobility by gel electrophoresis.

摘要

通过凝胶电泳以及与由两种特征明确的实验室毒株(Wa和DS-1)体外转录制备的探针杂交,对人轮状病毒的病毒RNA进行了比较。此外,还通过与由三种测试病毒制备的探针杂交来比较病毒RNA。在加拉加斯儿童医院急诊病房,于同一天从5至21个月大的婴幼儿中获取了13份标本(腹泻粪便),另外还从同一家医院6个月前获取了一份标本。当用溴化乙锭对电泳后的病毒RNA进行染色并通过紫外线检查时,根据RNA片段迁移率的差异区分出了五种不同的迁移模式(电泳型)。所采用的杂交技术仅允许对不同人轮状病毒的相应基因进行定性比较。其中10份标本含有足够的病毒,可产生用于杂交研究的足够RNA。通过杂交研究的8种病毒含有4至11个基因,这些基因与Wa探针特异性反应,产生迁移率与Wa病毒RNA或测试病毒RNA相似的双链RNA片段。另外两种病毒含有11个基因,这些基因与DS-1杂交探针特异性反应,产生迁移率与DS-1病毒RNA或测试病毒RNA相似的双链RNA片段。当从三种测试病毒制备杂交探针并用于比较不同的电泳型时,出现了更为复杂的情况。通过杂交研究时,迁移相似的相应基因不一定表现出同源性。此外,一些表现出同源性的相应基因在凝胶电泳中没有相同的迁移率。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2a22/347581/685975bd025d/iai00149-0260-a.jpg

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