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家蚕SOCS6的细胞定位及表达谱分析

发表时间:2014-09-15  浏览量:2149  下载量:1661
全部作者: 刘佳宾,曹广力,薛仁宇,胡小龙,彭景琨,贡成良
作者单位: 苏州大学基础医学与生物科学学院
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。研究克隆了家蚕 BmSOCS6基因的开放阅读框(open reading frame,ORF)序列,序列分析结果显示, BmSOCS6基因的ORF为918 bp,编码305个氨基酸,BmSOCS6蛋白具有典型的跨膜区和SH2超家族保守结构域。Real-time PCR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(+),在大肠杆菌(Escherichia 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: 分子生物学;SOSC6基因;表达谱;家蚕;细胞定位;免疫组化
Title: Cellular localization and expression profiling of SOCS6 in silkworm, Bombyx mori
Author: LIU Jiabin, CAO Guangli, XUE Renyu, HU Xiaolong, PENG Jingkun, GONG Chengliang
Organization: School of Biology & Basic Medical Sciences, Soochow University
Abstract: Suppressor of cytokine signaling (SOCS) gene is a negative regulator 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 encoded 305 amino acid residues, BmSOCS6 protein had a typical transmembrane region and a domain conserved in SH2 superfamily. The results of real-time PCR indicated that the abundance of BmSOCS6 mRNA in the testis was the highest, and 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 Escherichia 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 all expressed in germ cells.
Key words: molecular biology; SOSC6 gene; expression profile; Bombyx mori; cellular localization; immunohistochemistry
发表期数: 2014年9月第17期
引用格式: 刘佳宾,曹广力,薛仁宇,等. 家蚕SOCS6的细胞定位及表达谱分析[J]. 中国科技论文在线精品论文,2014,7(17):1730-1736.
 
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