克氏原螯虾CSGalNAcT-1基因功能初步研究

Preliminary study on function of CSGalNAcT-1 gene in red swamp crayfish,Procambarus clarkii

为探究硫酸软骨素N-乙酰半乳糖胺基转移酶-1(chondroitin sulfate N-acetylgalactosaminyltransferase-1,CSGalNAcT-1)在克氏原螯虾免疫反应中的作用,利用RACE技术成功获得克氏原螯虾CSGalNAcT-1基因完整cDNA序列,并通过荧光定量PCR分析该基因在各组织中的表达模式以及在CpG寡脱氧核苷酸(ODN)、嗜水气单胞菌和白斑综合征病毒(WSSV)刺激后的表达情况。结果显示,克氏原螯虾CSGalNAcT-1基因在中肠的相对表达量最高,其次是近胃段肠,在心脏、胃、肌肉和围食道神经中的相对表达量较低。克氏原螯虾肝胰腺、血细胞中CSGalNAcT-1基因表达量在嗜水气单胞菌和WSSV刺激下呈现出下调及上调,而在注射免疫刺激剂CpG ODN后,CSGalNAcT-1基因表达水平在大部分组织中呈现上调。表明克氏原螯虾CSGalNAcT-1基因可能在抵御外界病原侵染以及增强自身免疫反应的过程中发挥重要作用。

To investigate the role of chondroitin sulfate N-acetylgalactosaminyltransferase-1(CSGalNAcT-1)played in the immune response of Procambarus clarkii,in this study,the complete cDNA sequence of the P.clarkii CSGalNAcT-1 was obtained by RACE,and its expression patterns in response to stimulation of CpG oligodeoxynucleotide(ODN),Aeromonas hydrophila and white spot syndrome virus(WSSV)were investigated by real-time PCR.The results indicated that the relative expression of CSGalNAcT-1 was highest in the midgut,followed by the foregut,and less in the heart,stomach,muscle and periesophageal nerve in P.clarkii.Besides,the expression of CSGalNAcT-1 in the hepatopancreas and haemocytes which were considered as immune organs of crustacean,could be down-regulated and up-regulated by challenges of A.hydrophila and WSSV,respectively,and the expression was up-regulated in most of the tissues after injected with CpG ODN,indicating that the CSGalNAcT-1 gene may play an important role in resisting the pathogenic infection and enhancing the autoimmune response in P.clarkii.