Larsen M, Meinhardt F
Institut für Mikrobiologie, Universität Münster, Germany.
Curr Genet. 2000 Dec;38(5):271-5. doi: 10.1007/s002940000167.
The cytoplasmic linear plasmid pGKL2 of the yeast Kluyveromyces lactis was reported to harbour ten open reading frames (ORF1-ORF10). By re-examining the sequence, we identified an additional ORF encoding a polypeptide of 70 amino acids; and a homologous ORF with 70% similarity was also identified on plasmid pSKL of Saccharomyces kluyveri. As for pGKL2, the newly detected ORF11 is located at the same position with the same direction. ORF11 transcripts of pGKL2 were verified by reverse transcriptase-polymerase chain reaction; and the corresponding promoter activity was proven by expression of a reporter gene driven by the ORF11 upstream conserved sequence, indicating that ORF11 constitutes a functional gene.
据报道,乳酸克鲁维酵母的细胞质线性质粒pGKL2含有十个开放阅读框(ORF1 - ORF10)。通过重新检查该序列,我们鉴定出一个额外的编码70个氨基酸多肽的开放阅读框;并且在克鲁维酵母的质粒pSKL上也鉴定出一个相似度为70%的同源开放阅读框。至于pGKL2,新检测到的ORF11位于相同位置且方向相同。通过逆转录 - 聚合酶链反应验证了pGKL2的ORF11转录本;并且通过由ORF11上游保守序列驱动的报告基因的表达证明了相应的启动子活性,表明ORF11构成一个功能基因。