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糖胺聚糖在抗病毒、消炎和抗氧化等免疫反应发挥重要作用。磺基转移酶(sulfotransferase)是糖胺聚糖生物合成的关键酶,Carbohydrate sulfotransferase 9(CHST9)可以将磺酸基从3’-磷酸腺苷酰-5’磷酸硫酸盐(PAPS)转移到半乳糖残基非还原末端的4位碳上。为研究CHST9在马氏珠母贝免疫调节中的作用,本研究克隆得到马氏珠母贝CHST9(Pinctada martensii CHST9,Pm CHST9)c DNA全长序列并检测其在不同组织中的表达模式。利用RACE技术,得出Pm CHST9序列全长1 388 bp,其中5’UTR为122 bp,3’UTR为192 bp,开放阅读框(ORF)为1 074 bp,编码357个氨基酸;Pm CHST9氨基酸序列分子量为42 889.2 Da,等电点为9.28,脂溶性系数为81.32,总平均亲水性为-0.493;氨基酸序列分析得出Pm CHST9具有典型的磺基转移酶-2结构域和一个跨膜结构域;多序列比对结果显示Pm CHST9与其它物种的CHST9同源性较低;荧光定量PCR检测结果表明Pm CHST9在鳃中表达量最高,其次是足和肝胰腺。综上所述,Pm CHST9可能参与马氏珠母贝的免疫调节作用,为进一步阐述Pm CHST9在马氏珠母贝中的免疫机制提供基础资料。
Glycosaminoglycans play an important role in antiviral, anti-inflammatory and anti-oxidant immune responses. Sulfotransferases are key enzymes in the glycosaminoglycan biosynthesis and Carbohydrate sulfotransferase 9 (CHST9) can transfer sulfonate groups from 3’-phospho-adenylate-5’-phosphate sulfate (PAPS) to galactose Residue non-reducing end of the carbon at the 4 position. In order to study the role of CHST9 in immune regulation of Pinctada martensii, we cloned the full-length cDNA of cST from Pinctada martensii CHST9 (Pm CHST9) and detected the expression pattern in different tissues. The full length of Pm CHST9 was 1 388 bp, which was 122 bp in 5’UTR, 192 bp in 3’UTR and 1 074 bp in open reading frame (ORF), encoding 357 amino acids. The amino acid sequence of Pm CHST9 The molecular weight was 42 889.2 Da, the isoelectric point was 9.28, the lipophilicity coefficient was 81.32, the total average hydrophilicity was -0.493. Amino acid sequence analysis showed that Pm CHST9 has a typical sulfotransferase-2 domain and a transmembrane structure Sequence analysis showed that Pm CHST9 shared less homology with CHST9 from other species. Fluorescent quantitative PCR showed that Pm CHST9 had the highest expression in the gill, followed by the foot and hepatopancreas. Taken together, Pm CHST9 may be involved in the immunomodulatory effects of Pseudomonas solanacearum, providing the basic information for further elucidating the immunological mechanism of Pm CHST9 in M. jeadense.