目的:克隆多疣壁虎细胞因子信号转导负调控蛋白3A(suppressor of cytokine signaling,SOCS3A)基因的c DNA全长序列,并分析其结构特征;检测SOCS3A蛋白在多疣壁虎断尾损伤脊髓中的表达变化。方法:利用c DNA末端快速扩增(rapid amplification of c DNA ends,RACE)技术克隆得到多疣壁虎SOCS3A的c DNA全长序列;实时荧光定量聚合酶链反应(real-time polymerase chain reaction,RT-PCR)方法检测多疣壁虎断尾脊髓中SOCS3A的表达。结果:克隆得到多疣壁虎SOCS3A的c DNA全长序列。该序列全长1 152 bp,编码202个氨基酸,与爬行动物SOCS3同源性高度保守;多疣壁虎断尾损伤后,SOCS3A在断尾脊髓中的表达先下调随后上调。结论:成功克隆了多疣壁虎SOCS3A的c DNA全长序列,共1 152 bp,编码202个氨基酸。SOCS3A在多疣壁虎断尾再生脊髓中表达升高。
Objective: To clone the full length cDNA of suppressor of cytokine signaling 3A(SOCS3A) from Gekko japonicus, and characterize its primary structure; then we will examine its expression changes in the spinal cord following gecko tail amputation. Methods: To obtain the full-length of SOCS3A cDNA from gecko by rapid amplification of cDNA ends(RACE) technique. The expression changes of SOCS3 in the spinal cord were analyzed by real-time polymerase chain reaction (RT- PCR). Results: The SOCS3A cDNA of gecko was cloned with the full length of 1 152 bp encoding a protein of 202 amino acids. SOCS3A showed a high conservation during the phylogeny, especially among reptiles. The expression of SOCS3A was upregulated in the regenerating spinal cord following gecko tail amputation. Conclusion: The SOCS3A cDNA of gecko was cloned with the full length of 1 152 bp encoding a protein of 202 amino acids. The expression of SOCS3A was upregulated in the regenerating spinal cord following gecko tail amputation.