克氏原螯虾TUSC3基因的鉴定与免疫功能分析

Characterization and Immunological Function of Tumor Suppressor Candidate 3 (TUSC3) in Crayfish Procambarus clarkii

为了探究TUSC3 (Tumor suppressor candidate 3)基因在克氏原螯虾中的生物学功能,本研究利用PCR法克隆获得了该基因序列,并采用实时荧光定量PCR技术分析了该基因在克氏原鳌虾不同组织中的表达特征以及免疫诱导物(LPS和Poly I:C)对肠、腮和血淋巴细胞中TUSC3基因表达水平的影响。结果表明TUSC3基因编码326个氨基酸,预测蛋白与南美白对虾的TUSC3蛋白同源性较高(93.25%),包含一个保守的OST3/OST6同源功能区。PCR检测结果显示该基因在被检各组织中均有表达,且在血细胞组织中表达量最高,心脏组织中最低。LPS和Poly I:C刺激后,TUSC3基因的表达量总体表现为上升趋势,但不同组织及不同诱导物之间存在差异。这些结果表明TUSC3基因在克氏原螯虾的免疫反应过程中具有一定功能。

In order to explore the biological function of tumor suppressor candidate 3 in Procambarus clarkii, a TUSC3 gene was cloned using PCR and real-time fluorescent quantitative PCR method was used to detect the expression patterns of TUSC3 in various tissues of P. clarkii as well as the effect of immune inducer LPS and Poly I:C on the TUSC3 expression level in gut, gill and hemocytes. This gene encoded 326 amino acids with a conserved OST3/OST6 domain and the predicted protein was highly homologous to that in Penaeus vannamei (93.25%). PCR test results show TUSC3 was extensively expressed in all examined tissues with the highest expression level in hemocytes and lowest in heart. The expression level of TUSC3 was significantly increased after lipopolysaccharide (LPS) or polyinosinic-polycytidylic acid (poly I:C) induction. However, the expression patterns of TUSC3 differed in various tissues. These results suggested that TUSC3 played a role in immune response in P. clarkii.