Boniotto Michele, Ventura Mario, Cardone Maria Francesca, Boaretto Francesca, Archidiacono Nicoletta, Rocchi Mariano, Crovella Sergio
Università di Trieste, Dipartimento di Scienze della Riproduzione e dello Sviluppo, Italy.
Genome. 2002 Oct;45(5):973-6. doi: 10.1139/g02-048.
We have isolated and cloned an 800-bp highly repeated DNA (HRDNA) sequence from Lemur catta (LCA) and described its localization on LCA chromosomes. Lemur catta HRDNA sequences were localized by performing FISH experiments on standard and elongated metaphasic chromosomes using an LCA HRDNA probe (LCASAT). A complex hybridization pattern was detected. A strong pericentromeric hybridization signal was observed on most LCA chromosomes. Chromosomes 7 and 13 were lit in pericentromeric regions, as well as in the interspersed heterochromatin. Chromosomes 1, 3, 4, 17, 19, X, and microchromosomes (20, 25, 26, and 27) showed no signals in the pericentromeric region, but chromosomes 3 and 4 showed a positive hybridization in heterochromatic regions. The 800-bp L catta HRDNA was species specific. We performed FISH experiments with the LCASAT probe on Eulemur macaco macaco (EMA) and Eulemur fulvus fulvus (EFU) metaphases and no positive signal of hybridization was detected. These findings were also confirmed by Southern blot analysis and PCR.
我们从环尾狐猴（LCA）中分离并克隆了一段800 bp的高度重复DNA（HRDNA）序列，并描述了其在LCA染色体上的定位。通过使用LCA HRDNA探针（LCASAT）对标准和伸长的中期染色体进行荧光原位杂交（FISH）实验，确定了环尾狐猴HRDNA序列的定位。检测到一种复杂的杂交模式。在大多数LCA染色体上观察到强烈的着丝粒周围杂交信号。7号和13号染色体在着丝粒周围区域以及散布的异染色质中被点亮。1、3、4、17、19、X号染色体和微染色体（20、25、26和27）在着丝粒周围区域没有信号，但3号和4号染色体在异染色质区域显示出阳性杂交。800 bp的环尾狐猴HRDNA具有物种特异性。我们用LCASAT探针对黑冠猕猴（EMA）和褐美狐猴（EFU）的中期染色体进行了FISH实验，未检测到阳性杂交信号。这些发现也通过Southern印迹分析和PCR得到了证实。
