Ronchetti E, Crovella S, Rumpler Y, Pellicciari C, Manfredi Romanini M G
Dipartimento di Biologia Animale, University of Pavia, Italy.
Cytogenet Cell Genet. 1993;63(1):1-5. doi: 10.1159/000133490.
The amounts of nuclear DNA and the AT and GC content of four Eulemur (Prosimii, Lemuridae) species and of an E. coronatus x E. macaco hybrid were measured by flow cytometry in peripheral blood leukocytes, following propidium iodide, Hoechst 33258, and mithramycin staining. Hoechst 33258 and mithramycin were also used to evaluate the base composition of genomic DNA in the chromosomes. The amount of DNA resisting C-banding pretreatment (C-heterochromatic DNA) was measured in metaphase chromosomes by static fluorometry. The genome of E. coronatus was significantly larger than the genomes of all other species examined, due to a higher content of pericentromeric, mainly GC-rich, heterochromatic DNA. The restriction banding patterns produced by BamHI digestion and ethidium bromide staining on extracted DNA were studied in the hybrid and its parental species (E. coronatus and E. macaco). The restriction banding pattern of the sole E. coronatus individual showed two bands which were repeated in the restriction banding pattern of the hybrid. The qualitative and quantitative differences of C-heterochromatic DNA in E. coronatus confirm the "splitting" processes and the phylogenetic relationships in the genus Eulemur suggested by Jung et al. (1992) on the basis of the restriction banding patterns produced by endonuclease digestion.
采用流式细胞术，对四种狐猴（原猴亚目，狐猴科）以及一只冠美狐猴（E. coronatus）与黑美狐猴（E. macaco）的杂交后代的外周血白细胞进行碘化丙啶、Hoechst 33258和光神霉素染色后，测量其核DNA含量以及AT和GC含量。Hoechst 33258和光神霉素还用于评估染色体中基因组DNA的碱基组成。通过静态荧光测定法在中期染色体中测量抗C带预处理的DNA量（C异染色质DNA）。由于着丝粒周围（主要富含GC）的异染色质DNA含量较高，冠美狐猴的基因组明显大于所有其他检测物种的基因组。对杂交后代及其亲本物种（冠美狐猴和黑美狐猴）提取的DNA进行BamHI酶切和溴化乙锭染色，研究其限制性酶切图谱。唯一的冠美狐猴个体的限制性酶切图谱显示有两条带，这两条带在杂交后代的限制性酶切图谱中重复出现。冠美狐猴中C异染色质DNA的定性和定量差异证实了Jung等人（1992年）基于内切酶消化产生的限制性酶切图谱所提出的狐猴属中的“分裂”过程和系统发育关系。
