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猫杯状病毒衣壳蛋白基因的核苷酸序列与表达

Nucleotide sequence and expression of the capsid protein gene of feline calicivirus.

作者信息

Neill J D, Reardon I M, Heinrikson R L

机构信息

National Animal Disease Center, U.S. Department of Agriculture, Ames, Iowa 50010.

出版信息

J Virol. 1991 Oct;65(10):5440-7. doi: 10.1128/JVI.65.10.5440-5447.1991.

Abstract

The sequence of the 3'-terminal 2,486 bases of the feline calicivirus (FCV) genome was determined. This region of the FCV genome, from which the 2.4-kb subgenomic RNA is derived, contained two open reading frames. The larger open reading frame, found in the 5' end of the subgenomic mRNA, contained 2,004 bases encoding a polypeptide of 73,467 Da. The smaller open reading frame, encoded in the 3' end of the mRNA, was composed of 318 bases, encoding a polypeptide of 12,185 Da. The AUG initiation codon of the second open reading frame overlapped the UGA termination codon of the first, with the sequence AUGA. The nucleotide sequence of the region containing this overlap resembles the -1 frameshift sequences of the retroviruses. The 5' end of the 2.4-kb subgenomic RNA was mapped by primer extension analysis. There were two apparent transcription initiation points, both of which were 5' to the AUG initiation codon of the large open reading frame. Transcription from these sites yielded RNA transcripts with 5' nontranslated leader regions of 17 and 18 bases. The total length of the 2.4-kb subgenomic RNA was 2,375 bases (from the 5'-most start site) excluding the poly(A) tail. Edman degradation of the purified capsid protein of FCV showed that the capsid protein was encoded by the large open reading frame. Western immunoblot analysis of FCV-infected cells using a feline anti-FCV antiserum demonstrated that translation of the capsid protein was detectable at 3 h postinfection and continued to accumulate until 8 h postinfection, the last time examined.

摘要

测定了猫杯状病毒(FCV)基因组3'末端2486个碱基的序列。FCV基因组的这一区域是2.4kb亚基因组RNA的来源,包含两个开放阅读框。在亚基因组mRNA的5'端发现的较大开放阅读框包含2004个碱基,编码一个73467Da的多肽。在mRNA的3'端编码的较小开放阅读框由318个碱基组成,编码一个12185Da的多肽。第二个开放阅读框的AUG起始密码子与第一个的UGA终止密码子重叠,序列为AUGA。包含这种重叠的区域的核苷酸序列类似于逆转录病毒的-1移码序列。通过引物延伸分析对2.4kb亚基因组RNA的5'端进行了定位。有两个明显的转录起始点,两者都在大开放阅读框的AUG起始密码子的5'端。从这些位点转录产生的RNA转录本具有17和18个碱基的5'非翻译前导区。2.4kb亚基因组RNA的总长度为2375个碱基(从5'最起始位点开始),不包括聚腺苷酸尾。对纯化的FCV衣壳蛋白进行的埃德曼降解表明,衣壳蛋白由大开放阅读框编码。使用猫抗FCV抗血清对FCV感染细胞进行的western免疫印迹分析表明,衣壳蛋白的翻译在感染后3小时可检测到,并持续积累直至感染后8小时,这是最后一次检测的时间。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b314/249032/c4535df5990d/jvirol00053-0328-a.jpg

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