Developmental and Evolutionary Analysis of Ancient Fish (Polypterus)

培养细胞系的建立：建立了塞内加尔多嘴鱼鳍、肾脏和胚胎的原代细胞培养。细胞培养初期，细胞生长良好。经多次传代后，大部分细胞迅速凋亡。在鳍、肾和胚胎中均观察到这种现象。虽然我们改变了培养基、胶原包衣皿等几种培养条件，但都没有成功。fosmid文库的构建与进化遗传分析：利用fosmid载体系统构建了多翅虫基因组文库。这个图书馆的独立殖民地会员超过50万。对Gcm2、Elovl5、Fbox93三个基因进行PCR筛选后，鉴定出单个Elovl5基因阳性克隆。这一结果表明，该fosmid文库的基因组覆盖率不够高，不足以分离出多翅虫基因组中的任意基因。

Establishment of cultured cell line : We have been established the primary cell culture from fin, kidney and embryos of Polypterus senegalus. Initial phase of cell culture, cells were growing well. After several passages, most of cells were rabidly dying by apoptosis. This phenomenon was observed in fins, kidney and embryo. Although we have changed several culture conditions such as medium and collagen coated dish, all efforts were not successful.Construction of fosmid library and evolutionary genetic analysis : We have constructed the Polypterus genomic library with fosmid vector system. Independent colony members of this library reached over 500,000. After PCR screening of three genes (Gcm2、Elovl5、Fbox93), single positive clone with the Elovl5 gene was identified. This result suggests the genome coverage of this fosmid library was not high enough for isolation of arbitral genes in Polypterus genome.