青蛤MAPK/p38基因的克隆及Poly∶Ic胁迫下的表达分析

目的]克隆青蛤MAPK/p38基因,并分析其在Poly:Ic胁迫下的表达情况。方法]利用转录组测序获得青蛤MAPK/p38基因的序列信息,采用生物信息学软件分析该基因在近缘生物间的进化关系;利用巢式及荧光定量PCR技术克隆青蛤MAPK/p38基因的结构域序列,并在Poly:Ic胁迫后立即分析该基因在青蛤各组织中的表达情况以及血淋巴中的时序性表达情况。结果]克隆得到青蛤MAPK/p38基因的结构域序列,分析发现该基因在物种进化中很保守,其在青蛤的血淋巴、肝脏、外套膜、闭壳肌和鳃中均有表达,其中在血淋巴中表达量最高,腮中表达量最低。Poly:Ic胁迫3 h后该基因的表达量达到最大值,与对照组差异极显著(P0.01)。结论]MAPK/p38基因所表达的产物是青蛤重要的免疫信号通路蛋白。

Cloning of MAPK/p38 in Cyclina sinensis and Its Expression under Poly∶Ic Strees

Objective] To clone the MAPK/P38 gene,and analyze its expression under Poly∶ Ic stress.Method] Using the transcrip tome sequencing won the Cyclina sinensis silk crack the original amp-activated protein kinase (MAPK/p38) gene sequence information,using bioinformatics software analyze the evolution of the relationship between the gene in the related biological.Using the nested and fluorescence quantitative PCR cloning technology and analyzing the MAPK/p38 gene expression in organizations,as well as Poly∶ Ic testing under the stress of the MAPK/p38 gene sequence in C.sinensis haemolymph expression.Result]The MAPK/p38 gene structural domain sequences was cloned,the analysis found that gene in evolution was very conservative,MAPK/p38 genes in C.sinensis haemolymph muscle,liver,coat film,closed shell and gills were expressed,which express in the haemolymph amount was highest,gills expressed in the lowest amount.The gene expression quantity under Ploy∶Ic stress in 3 h was the maximum,which had extremely significant difference with the control group (P＜ 0.01).Conclusion]The product expressed by the gene was important immune signaling pathways protein for C.sinesis.