Requena J M, Jimenez-Ruiz A, Soto M, Lopez M C, Alonso C
Centro de Biología Molecular, Universidad Autónoma de Madrid, Spain.
Mol Biochem Parasitol. 1992 Apr;51(2):271-80. doi: 10.1016/0166-6851(92)90077-w.
In the present paper we report the characterization of a repetitive DNA sequence from the genome of the Maracay strain of Trypanosoma cruzi. The sequence is 1025 nucleotides long and represent about 7% of the total nuclear DNA of the parasite with a copy number of 1-2 x 10(4) copies per genome. It is also present in a high copy number in several strains of T. cruzi but absent from other trypanosomatids and from the human genome. The DNA sequence is interspersed and present on many chromosomes. We show that the different copies of this repeat present extensive endonuclease restriction polymorphisms. Using 2 conserved oligonucleotide primers derived from the repetitive sequence, together with the polymerase chain reaction technique, the presence of less than 1/30 of a single parasite can be readily detected by ethidium bromide staining.
在本论文中,我们报道了来自克氏锥虫马拉开波菌株基因组的一个重复DNA序列的特征。该序列长度为1025个核苷酸,约占寄生虫总核DNA的7%,每个基因组的拷贝数为1 - 2×10⁴个。它在几种克氏锥虫菌株中也有高拷贝数存在,但在其他锥虫以及人类基因组中不存在。该DNA序列是散布的,存在于许多染色体上。我们表明,这个重复序列的不同拷贝呈现出广泛的内切酶限制性多态性。使用从该重复序列衍生的2个保守寡核苷酸引物,结合聚合酶链反应技术,通过溴化乙锭染色可以很容易地检测到单个寄生虫中少于1/30的量的存在。