论文部分内容阅读
Ras主要参与表皮生长因子家族调控细胞分裂的信号通路。采用末端快速扩增法获得了牙鲆(Paralichthys olivaceus)ras-2基因全长2 384 bp cDNA序列,该序列编码187个氨基酸,含有switchⅠ、switchⅡ、P-loop、CAAX box等Ras家族的结构特征,与川鲽(Platichthys flesus)ras-2氨基酸序列相似性最高(96.3%)。定量RT-PCR检测显示ras-2基因在牙鲆成体的心脏、肝脏、脾脏、肾脏、鳃、肠、肌肉、鳍条和皮肤组织均有不同程度的表达,其中鳍条表达量最高,肌肉的表达量最低;在牙鲆变态前和变态初相对表达量较高,随着变态进行,表达水平显著降低。牙鲆变态阶段的RNA整体原位杂交结果显示ras-2基因主要在鳍条、眼眶周围、颌、鳃、鼻孔、侧线等部位的表皮组织中表达,变态前在鳍褶和体侧线表达信号最强,变态阶段E期的信号稍微减弱,主要分布在支鳍骨和鳃,变态高峰期(F期和G期)信号主要集中在支鳍骨、鳃和颌,H期表达信号主要集中在鳃和支鳍骨。ras-2基因表达式型表明其可以通过调控细胞分裂的方式,参与牙鲆变态过程中眼睛移动、冠状幼鳍发育、侧线发育等多种器官的发育调控。
Ras is mainly involved in the epidermal growth factor family regulation of cell division of the signal pathway. The full length 2 384 bp cDNA sequence of ras-2 gene of Paralichthys olivaceus was obtained by rapid amplification of end sequences. The sequence encodes 187 amino acids and contains the structural characteristics of the Ras family of switchⅠ, switchⅡ, P-loop and CAAX box , And ras-2 had the highest similarity with that of Platichthys flesus (96.3%). Quantitative RT-PCR results showed that ras-2 gene was expressed in different degrees in heart, liver, spleen, kidney, gill, intestine, muscle, ray and skin tissues of adult flounder, with the highest expression of muscular The lowest expression level was observed in the early stage of metamorphosis and metamorphosis of the Japanese flounder. With the metamorphosis, the expression level decreased significantly. The whole-mount RNA in situ hybridization of Japanese flounder metamorphosis showed that the ras-2 gene was mainly expressed in the epidermis of the rays, eyes, periorbital, gills, nostrils and lateral lines. Before the metamorphosis, The signal of the E and E phase of the strong and abnormal stage was slightly weakened, mainly distributed in the branches and gills of the fin, and the signals in the abnormal period (F and G) were mainly in the fins, gills and jaws. The signals in the H phase mainly concentrated in the gills And support fin bone. The ras-2 gene expression pattern indicates that it can regulate the development of multiple organs such as eye movement, coronoid fimbriation and lateral line development by regulating cell division.