Laboratory of Animal Cytogenetics and Comparative Genomics (ACCG), Department of Genetics, Faculty of Science, Kasetsart University, Bangkok, 10900, Thailand.
Interdisciplinary Program in Genetic Engineering, Graduate School, Kasetsart University, Bangkok, 10903, Thailand.
Sci Rep. 2019 Oct 29;9(1):15459. doi: 10.1038/s41598-019-51863-w.
To better understand PBI-DdeI satellite DNA located in the centromeric region of python, molecular evolution analysis was conducted on 40 snake species. A ladder-like pattern of DNA bands with repetition of the 194-210 bp monomer was observed in 15 species using PCR. Molecular cloning was performed to obtain 97 AT-rich monomer sequences. Phylogenetic and network analyses showed three PBI-DdeI subfamilies with sequences grouped in species-specific clusters, suggesting rapid evolution. Slow evolution was found in eight species with shared PBI-DdeI sequences, suggesting recent species diversification, allowing PBI-DdeI no time to diverge, with limited homogenization and fixation processes. Quantitative real-time PCR showed large differences in copy number between Python bivittatus and other snakes, consistent with repeat scanning of whole genome sequences. Copy numbers were significantly higher in female Naja kaouthia than in males, concurring with chromosomal distribution of PBI-DdeI specifically localized to female W chromosomes. PBI-DdeI might act as an evolutionary driver with several repeats to promote W chromosome differentiation and heterochromatinization in N. kaouthia. Analysis revealed PBI-DdeI with a reduced copy number, compared to P. bivittatus, in most snakes studied, and it is possible that it subsequently dispersed and amplified on W chromosomes with different functional roles in N. kaouthia.
为了更好地了解位于巨蟒着丝粒区的 PBI-DdeI 卫星 DNA,对 40 种蛇进行了分子进化分析。通过 PCR,在 15 种蛇中观察到具有重复 194-210bp 单体的梯状 DNA 带模式。通过分子克隆获得了 97 个富含 AT 的单体序列。系统发育和网络分析显示,有三个 PBI-DdeI 亚家族,序列在种内聚类,表明快速进化。在 8 个具有共享 PBI-DdeI 序列的物种中发现了缓慢进化,表明最近的物种多样化,使 PBI-DdeI 没有时间分化,有限的同质化和固定化过程。实时定量 PCR 显示,球蟒和其他蛇之间的拷贝数存在很大差异,与全基因组序列的重复扫描一致。Naja kaouthia 雌性的拷贝数明显高于雄性,与 PBI-DdeI 特异性定位于雌性 W 染色体的染色体分布一致。PBI-DdeI 可能作为一个进化驱动因子,具有多个重复,促进 N. kaouthia 的 W 染色体分化和异染色质化。分析显示,与球蟒相比,大多数研究的蛇中 PBI-DdeI 的拷贝数减少,它可能随后在 N. kaouthia 的 W 染色体上分散和扩增,具有不同的功能作用。