家蚕SOCS6的细胞定位及表达谱分析.docVIP

  家蚕 SOCS6 的细胞定位及表达谱分析# 刘佳宾，曹广力，薛仁宇，胡小龙，彭景琨，贡成良** （苏州大学基础医学与生物科学学院，苏州 215123） 5 10 15 20 25 30 摘要：细胞信号转导抑制因子（suppressor of cytokine signaling，SOCS）基因是 JAK/STAT 信号通路的负调节基因。本研究克隆了家蚕 BmSOCS6 基因的 ORF 序列，序列分析结果显 示，BmSOCS6 基因的 ORF 为 918 bp，编码 305 aa，BmSOCS6 蛋白具有典型的跨膜区和 SH2 超家族保守结构域。qPCR 分析结果显示，BmSOCS6 mRNA 丰度在家蚕精巢中最高，在中 肠、脂肪、血细胞和丝腺相对较低。将 BmSOCS6 基因克隆进原核表达载体 pET28a(+)，在 E. coli 中表达了重组 BmSOCS6。利用重组蛋白免疫小鼠，制作了抗 BmSOCS6 多抗。细胞 定位结果显示 BmSOCS6 主要定位在 BmN 细胞的细胞质中，免疫荧光研究结果显示， BmSOCS6 在生殖细胞中均有表达。 关键词：家蚕；SOSC6 基因；表达谱；细胞定位，免疫组化 中图分类号：S881.2；Q751 Cellular localization and expression profiling of SOCS6 in silkworm, Bombyx mori LIU Jiabin, CAO Guangli, XUE Renyu, HU Xiaolong, PENG Jingkun, GONG Chengliang (School of Biology Basic Medical Science, Soochow University, Suzhou 215123) Abstract: Suppressor of cytokine signaling (SOCS) gene is a negetive regulate gene of JAK/STAT signal pathway. In this study, the open reading frame (ORF) of silkworm BmSOCS6 was cloned. Sequence analysis showed that the ORF was 918 bp in size and encodes 305 amino acid residues, BmSOCS6 protein has a transmembrane region and a typical domain conserved in SH2 superfamily. The results of qPCR indicated that the abundance of BmSOCS6 mRNA in the testis was the highest, was lower in the midgut, fat body, hemocyte and silk gland.The BmSOCS6 gene was cloned into the prokaryotic expression vector pET28a (+), recombinant BmSOCS6 was expressed in E. coli, the recombinant BmSOCS6 was used to immunize mice to generate anti-BmSOCS6 polyclonal antibody. Cellular localization showed BmSOCS6 mainly localized in the cytoplasm of BmN cells, immunofluorescence studies showed that BmSOCS6 were both expressed in germ cells. Key words: Bombyx mori; SOSC6 gene; expression profile; cellular localization; Immunohistochemistry 35 0 引言 SOCS 基因是 JAK/STAT 信号通路的负调节基因。SOCS 家族包括八种蛋白：CIS （Cytokine-inducible SH2-containing protein）和 SOCS1-SOCS7。CIS 和 SOCS1-SOCS7 都包 含一个 SH2 中心域，一个作为 SOCS box 的保守的 C 末端和一个特异性的 N 末端。SOCS 成 40 员集中分布在细胞质