Fu G, Lambson B, Barker D
Department of Pathology, University of Cambridge, UK.
FEMS Microbiol Lett. 1999 Mar 1;172(1):65-71. doi: 10.1111/j.1574-6968.1999.tb13451.x.
In this study, we have sequenced more than 100 clones of minicircle DNA from Herpetomonas samuelpessoai. An unusual amplification approach was developed to amplify minicircle DNA by using a pair of complementary primers designed from a universal stretch of minicircle sequence. Sequence analysis shows that the kinetoplast minicircles in Herpetomonas with a size of 1.3 kb are organised into two conserved regions and two variable regions which are located 180 degrees apart. The potential gRNA genes are encoded in variable regions of minicircle approximately 360 bp from CSB-3 (conserved sequence block 3). A conserved upstream sequence located 30 nt before the gRNA genes was identified and is related to the gRNA genes in sequence organisation. A potential role(s) of this sequence in gRNA transcription is discussed.
在本研究中,我们对来自塞缪尔佩索阿动基体虫(Herpetomonas samuelpessoai)的100多个微小环DNA克隆进行了测序。我们开发了一种不同寻常的扩增方法,通过使用从微小环序列的一段通用序列设计的一对互补引物来扩增微小环DNA。序列分析表明,动基体虫中大小为1.3 kb的动基体微小环被组织成两个保守区域和两个可变区域,这两个可变区域相隔180度。潜在的引导RNA(gRNA)基因编码在微小环的可变区域,距离保守序列块3(CSB-3)约360 bp。在gRNA基因之前30 nt处鉴定出一个保守的上游序列,该序列在序列组织上与gRNA基因相关。本文讨论了该序列在gRNA转录中的潜在作用。