黄鳝Nup93基因的分子克隆及其在性腺和肾的显著表达

核孔蛋白（Nucleoporins，Nups）是核孔复合体（Nuclear pore complexes，NPC）的重要组成成分，核孔复合体可以控制细胞内信号分子在核质问的双向转运，从而控制基因表达、细胞增殖和分化。在构建的黄鳝精巢SMART cDNA文库中，采用差异筛选的方法得到黄鳝核孔蛋白家族中Nup93基因的3’端片段，根据此段序列设计引物，使用兼并PCR和5＇RACE方法克隆得到此基因的全长cDNA。序列比对显示该基因与酵母Nic96、斑马鱼Nup93和人类Nup93的同源性分别为36．5％、94．6％和90．5％。进化树分析显示，黄鳝Nup93与其他鱼类的Nup93归为一支。采用荧光定量PCR方法对不同性别黄鳝的性腺和其他组织内该基因的表达作定量分析发现，Nup93在性腺和肾中的表达量远高于其他组织，而且表达量存在一定的性别差异。这一结果提示Nup93可能与性腺发育相关。

Molecular Cloning of the Rice Field Eel Nup93 with Predominant Expression in Gonad and Kidney

Nucleoporins (Nups) are important components of nuclear pore complexes (NPCs).NPCs control gene expression,cells proliferation and differentiation by mediating exchange of cellular signal molecules on both nuclear and cytoplasmic sides.Using subtractive screening,3'end fragment of Nup93 from the testis cDNA library of the rice field eel was obtained.Full-length cDNA of the gene was further cloned by degenerate PCR and 5'RACE methods.Sequence analysis indicated that the homology of the rice field eel Nup93 were 36.5% with yeast Nic96,94.6% and 90.5% with Nup93 of zebrafish and human,respectively.Phylogenetic analysis showed that the rice field eel Nup93 fits with Nup93 of the other fishes.Real-time PCR result showed that expression of Nup93 in gonads and kidney were much higher than in other tissues,and different expression quantities among gonads of three sexes were also observed,suggesting that Nup93 may involve in gonad development.