COMPLETE NUCLEOTIDE-SEQUENCE OF AN ATTENUATED HEPATITIS-A VIRUS - COMPARISON WITH WILD-TYPE VIRUS
COMPLETE NUCLEOTIDE-SEQUENCE OF AN ATTENUATED HEPATITIS-A VIRUS - COMPARISON WITH WILD-TYPE VIRUS
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DOI：
10.1073/pnas.84.8.2497
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发表时间：
1987-04-01
影响因子：
11.1
通讯作者：
PURCELL, RH
中科院分区：
文献类型：
--
作者：
COHEN, JI;ROSENBLUM, B;PURCELL, RH
减毒甲型肝炎病毒的完整核苷酸序列。HAV HM-175/7 MK-5，由克隆的cDNA确定。该病毒来源于野生型HAV HM-175，在非洲绿色猴肾细胞中传代32次。所得的细胞培养适应病毒对黑猩猩是减毒的。将该病毒在猴肾细胞中再传代3次，以获得足够的病毒用于分子克隆，并将其命名为HM-175/7 MK-5。获得了三个重叠的cDNA克隆，它们一起跨越整个基因组。比较野生型病毒（在绒猴肝脏中体内繁殖）与减毒病毒（在细胞培养物中生长）的cDNA的核苷酸序列，显示分布在整个基因组中的24个核苷酸变化。5个碱基缺失发生在5“非编码区，编码区16个碱基替换中有12个导致氨基酸改变。氨基酸的变化发生在病毒衣壳蛋白VP 1和VP 2和几个非结构蛋白。因此，少量的核苷酸变化是适应细胞培养和HAV株HM-175减毒的原因。
The complete nucleotide sequence of an attenuated hepatitis A virus. HAV HM-175/7 MK-5, was determined from cloned cDNA. This virus was derived from wild-type HAV HM-175 after 32 passages in African green monkey kidney cells. The resultant cell culture-adapted virus is attenuated for chimpanzees. This virus was passaged an additional three times in monkey kidney cells to obtain sufficient virus for molecular cloning and was designated HM-175/7 MK-5. Three overlapping cDNA clones were obtained that together spanned the entire genome. Comparison of the nucleotide sequence of cDNA from wild-type virus (propagated in marmoset liver in vivo) with attenuated virus (grown in cell culture) showed 24 nucleotide changes distributed throughout the genome. Five base deletions occurred in the 5'' noncoding region, and 12 of the 16 base substitutions in the coding region resulted in amino acid changes. Amino acid changes occurred in viral capsid proteins VP1 and VP2 and several of the nonstructural proteins. Thus, a small number of nucleotide changes are responsible for adaptation to cell culture and attenutation of HAV stain HM-175.