脊髓灰质炎病毒cDNA的分子克隆及病毒基因组完整核苷酸序列的测定。
Molecular cloning of poliovirus cDNA and determination of the complete nucleotide sequence of the viral genome.
作者信息
Racaniello V R, Baltimore D
出版信息
Proc Natl Acad Sci U S A. 1981 Aug;78(8):4887-91. doi: 10.1073/pnas.78.8.4887.
Abstract
The complete 7410 nucleotide sequence of poliovirus type I genome was obtained from cloned cDNA. Double-stranded poliovirus cDNA was synthesized and inserted into the Pst I site of plasmid pBR322, and three clones were derived that together provided DNA copies of the entire poliovirus genome. Two of the clones contained inserts of 2.5 and 6.5 kilobases and represented all but the 5' 115 bases of poliovirus RNA. A third clone was generated from primer-extended DNA and contained sequences from the 5' end of the viral RNA. An open reading frame that was identified in the nucleotide sequence starting 743 bases from the 5' end of the RNA and extending to a termination codon 71 bases from the 3' end contained known poliovirus polypeptide sequence.
摘要
I型脊髓灰质炎病毒基因组完整的7410个核苷酸序列是从克隆的cDNA中获得的。合成了双链脊髓灰质炎病毒cDNA，并将其插入质粒pBR322的Pst I位点，得到了三个克隆，它们共同提供了整个脊髓灰质炎病毒基因组的DNA拷贝。其中两个克隆含有2.5和6.5千碱基的插入片段，代表了脊髓灰质炎病毒RNA除5'端115个碱基以外的所有序列。第三个克隆是由引物延伸DNA产生的，包含病毒RNA 5'端的序列。从RNA 5'端起始743个碱基处开始并延伸至3'端71个碱基处的终止密码子的核苷酸序列中鉴定出一个开放阅读框，其包含已知的脊髓灰质炎病毒多肽序列。
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