Charting latency transcripts in Kaposi's sarcoma-associated herpesvirus by whole-genome real-time quantitative PCR
Charting latency transcripts in Kaposi's sarcoma-associated herpesvirus by whole-genome real-time quantitative PCR
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DOI：
10.1128/jvi.76.12.6213-6223.2002
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发表时间：
2002-06-01
影响因子：
5.4
通讯作者：
Dittmer, DP
中科院分区：
文献类型：
--
作者：
Fakhari, FD;Dittmer, DP
潜伏或裂解生活周期的划分是所有疱疹病毒科的基础。在卡波西肉瘤相关疱疹病毒（KSHV）（人类疱疹病毒8）的情况下，潜伏基因已经涉及细胞自主转化，而某些裂解基因通过旁分泌机制获得肿瘤友好环境。为了查询KSHV转录，我们设计并验证了一个高通量，高特异性，高灵敏度，实时定量逆转录PCR阵列。这种新方法适用于许多人类病原体。它的首次使用表明，KSHV LAMA，v-cyclin和v-FLIP的mRNA水平在病毒再活化后的任何时间都不会增加。拉娜-2/vIRF-3的mRNA对病毒再活化具有类似的抗性。相反，其他所有潜在的或裂解的信息都被诱导了。因此，拉娜、v-FLIP、v-cyclin和拉娜-2构成KSHV基因组中一组独特调节的转录物。
The division into a latent or lytic life cycle is fundamental to all herpesviridae. In the case of Kaposi's sarcoma-associated herpesvirus (KSHV) (human herpesvirus 8), latent genes have been implicated in cell autonomous transformation, while certain lytic genes procure a tumor friendly milieu through paracrine mechanism. To query KSHV transcription, we devised and validated a high-throughput, high-specificity, high-sensitivity, real-time quantitative reverse transcription-PCR array. This novel methodology is applicable to many human pathogens. Its first use demonstrated that the mRNA levels for KSHV LAMA, v-cyclin, and v-FLIP do not increase at any time after viral reactivation. The mRNA for LANA-2/vIRF-3 is similarly resistant to viral reactivation. In contrast, every other latent or lytic message was induced. Hence, LANA, v-FLIP, v-cyclin, and LANA-2 constitute a group of uniquely regulated transcripts in the KSHV genome.