Comparative genome maps of the pangolin, hedgehog, sloth, anteater and human revealed by cross-species chromosome painting: further insight into the ancestral karyotype and genome evolution of eutherian mammals

To better understand the evolution of genome organization of eutherian mammals, comparative maps based on chromosome painting have been constructed between human and representative species of three eutherian orders: Xenarthra, Pholidota, and Eulipotyphla,

GENETIC DIVERSITY IN THE CHINESE PANGOLIN (MANIS-PENTADACTYLA) INFERRED FROM PROTEIN ELECTROPHORESIS

We examined protein polymorphism of Chinese pangolins (Manis pentadactyla) from Yunnan Province of China, including two forms of three brown and nine dusky Chinese pangolins. Sixty-two genetic loci were screened; 12 loci were found to be polymorphic. The percentage of polymorphic loci (P) is 0.194, the mean individual heterozygosity (H) is 0.078, and the mean number of alleles (A) is 1.258. Furthermore, we calculated the genetic distance (D) between the two forms and found a low level of genetic divergence (D = 0.0206) between them, which indicates an almost-indistinguishable divergence at the level of proteins.

Chromosomal rearrangements underlying karyotype differences between Chinese pangolin (Manis pentadactyla) and Malayan pangolin (Manis javanica) revealed by chromosome painting

The Chinese pangolin (Manis pentadactyla), a representative species of the order Pholidota, has been enlisted in the mammalian whole-genome sequencing project mainly because of its phylogenetic importance. Previous studies showed that the diploid number o

GENETIC DIVERSITY IN THE CHINESE PANGOLIN (MANIS-PENTADACTYLA) - INFERRED FROM RESTRICTION ENZYME ANALYSIS OF MITOCHONDRIAL DNAS

Two different forms of Chinese pangolins can be recognized according to the color of their scales, i.e., brown and dusky. We analyzed mitochondrial DNA (mtDNA) purified from the livers of seven dusky and six brown Chinese pangolins from the same locality, using cleavage patterns from 19 restriction enzymes. From the 19 6-bp recognition enzymes used, 51-56 sites were observed. By combining the cleavage patterns for each enzyme, the 13 samples were classified into four restriction types: two in dusky and two in brown Chinese pangolins. The estimated number of nucleotide substitutions per site in dusky and brown types is 0.002, and that between dusky and brown types is 0.012. Divergence between brown and dusky forms began 0.6 Myr ago, provided the mean rate of sequence divergence is 0.02 per Myr in mtDNA. Our results suggest that there is considerable divergence in Chinese pangolins, and brown and dusky Chinese pangolins may be quite different forms or, at least, belong to different maternal groups.

中国穿山甲细菌人工染色体文库的构建及其特征鉴定

鳞甲目 （Pholidota）作为真兽亚纲18个现生目之一，是理解哺乳动物系统分类和进化的一个关键类群。穿山甲的形态十分特殊，它的系统发生地位历来就是人们关注的对象，搞清穿山甲的系统发生地位将有助于理解和重建整个哺乳动物的系统发生过程。目前，中国穿山甲已被列入“哺乳动物基因组计划”24个待测序物种其中之一。细胞遗传学的研究表明中国穿山甲的核型具有多态性。 细菌人工染色体 （BAC）文库目前已成为基因组学研究领域的最为重要的研究工具之一。各种动植物细菌人工染色体文库的逐渐丰富为后基因组时代比较基因组研究奠定了良好的基础。中国穿山甲基因组DNA文库的构建将有力地促进穿山甲染色体演化、哺乳动物比较基因组学及其系统发育学的研究。 本研究在世界上首次成功地构建了一个具有约8倍基因组覆盖率的中国穿山甲细菌人工染色体文库。该基因组文库含有208,512个克隆，储存在543块384孔板中。对随机选取的171个克隆的Not1酶切和脉冲场电泳结果分析显示: 64.5%的克隆插入片段大小介于100-149 Kb之间， 75.6%的克隆插入片段大于100kb。平均插入片段大小为111.4Kb; 非重组克隆 （无插入片段）的比率为9.9%。同时，我们通过荧光原位杂交把105个BAC克隆定位到了中国穿山甲染色体上的特异区域, 构建了中国穿山甲的BAC细胞遗传学图谱。初步的染色体定位结果表明，BAC克隆在基因组中的分布没有偏向性。 中国穿山甲BAC文库的成功构建，丰富了哺乳动物基因组文库资源。该文库除有利于比较基因组研究之外，也为深化保护遗传学研究奠定了基础。BAC细胞遗传学图谱的构建，将为中国穿山甲全基因组测序中产生的序列重叠群在染色体上的定向和定位奠定基础。