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人巨细胞病毒基因组在有效感染细胞中的转录。

Transcription of the human cytomegalovirus genome in productively infected cells.

作者信息

Chua C C, Carter T H, St Jeor S

出版信息

J Gen Virol. 1981 Sep;56(Pt 1):1-11. doi: 10.1099/0022-1317-56-1-1.

Abstract

Nuclear and cytoplasmic RNAs, synthesized in cells productively infected with human cytomegalovirus (HCMV) were analysed at various times after infection by liquid and filter DNA-RNA hybridization. Results of these experiments have revealed that: (i) the fraction of the genome transcribed increased as infection progressed. In the nucleus, transcripts represented approx. 20% of the virus DNA sequences at both 2 and 4 h post-infection (p.i.) and 36% of the virus DNA at 40 h p.i; (ii) the increase in virus sequences among nuclear transcripts at late times was prevented by the DNA synthesis inhibitor, 2'-deoxyfluorouridine; (iii) early virus RNA transcripts were subset of those represented in late RNA; (iv) two classes of early RNA were identified by competition hybridization; (v) approx. 10% of the late nuclear transcripts were symmetrical. Results of filter hybridization at DNA excess indicated that virus-specific RNA represented 0.6% of RNA labelled from 0 to 2 h p.i., and 1.8% of RNA labelled from 28 to 30 h. Polyadenylated RNA isolated from cytoplasm represented 1.2% and 10% of labelled mRNA at 2 h and 30 h respectively. Our data show that during productive infection of human cells by HCMV, gene expression is under temporal, quantitative and post-transcriptional control.

摘要

通过液体和滤膜DNA - RNA杂交技术,对在人巨细胞病毒(HCMV)有效感染的细胞中合成的核RNA和胞质RNA在感染后的不同时间进行了分析。这些实验结果表明:(i)随着感染的进展,转录的基因组部分增加。在细胞核中,感染后2小时和4小时,转录本约占病毒DNA序列的20%,感染后40小时占病毒DNA的36%;(ii)DNA合成抑制剂2'-脱氧氟尿苷可阻止后期核转录本中病毒序列的增加;(iii)早期病毒RNA转录本是晚期RNA中所代表转录本的一个子集;(iv)通过竞争杂交鉴定出两类早期RNA;(v)约10%的晚期核转录本是对称的。DNA过量时的滤膜杂交结果表明,病毒特异性RNA在感染后0至2小时标记的RNA中占0.6%,在感染后28至30小时标记的RNA中占1.8%。从细胞质中分离的聚腺苷酸化RNA在2小时和30小时分别占标记mRNA的1.2%和10%。我们的数据表明,在HCMV对人细胞的有效感染过程中,基因表达受到时间、数量和转录后控制。

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