Mankertz J, Buhk H J, Blaess G, Mankertz A
Fachbereich Gentechnik und Genetik, Robert Koch-Institut, Berlin, Germany.
Virus Genes. 1998;16(3):267-76. doi: 10.1023/a:1008022521329.
This study focuses on gene expression of porcine circovirus (PCV) in order to identify viral genes and their corresponding mRNA transcripts. By northern blot analysis, the existence of three mRNAs could be demonstrated. Two mRNAs are encoded by the viral (-)-strand and one is encoded by the viral (+)-strand. The (+)-strand encoded mRNA transcript is 990 nucleotides (nt) long and corresponds to the open reading frame (ORF) 1, as shown by S1 mapping. The start point of this transcript is located at pos. 1238, as determined by primer extension analysis and rapid amplification of cDNA ends (RACE). The transcript is spliced as shown by direct reverse sequencing and RACE. It contains an untranslated "leader"-sequence 119 nt in size (pos. 1238 to 1120) which is joined to exon 2 of the ORF 1 transcript at pos. 737. The transcriptional regulatory elements have been identified functionally by CAT assays. They are located within a 258 base points (bp) fragment (pos. 1168 to 1425).
本研究聚焦于猪圆环病毒（PCV）的基因表达，以鉴定病毒基因及其相应的mRNA转录本。通过Northern印迹分析，可证实存在三种mRNA。两种mRNA由病毒（-）链编码，一种由病毒（+）链编码。如S1作图所示，由（+）链编码的mRNA转录本长度为990个核苷酸（nt），对应于开放阅读框（ORF）1。通过引物延伸分析和cDNA末端快速扩增（RACE）确定，该转录本的起始点位于第1238位。如直接反向测序和RACE所示，该转录本发生了剪接。它包含一个大小为119 nt的非翻译“前导”序列（第1238位至1120位），该序列在第737位与ORF 1转录本的外显子2相连。通过CAT分析在功能上鉴定了转录调控元件。它们位于一个258个碱基对（bp）的片段（第1168位至1425位）内。
