盐藻蛋白酶体亚基PSMD7在鞭毛解聚后的表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2017.03.014

生物技术通报 ›› 2017, Vol. 33 ›› Issue (3): 93-99.doi: 10.13560/j.cnki.biotech.bull.1985.2017.03.014

盐藻蛋白酶体亚基PSMD7在鞭毛解聚后的表达分析

石科^1,2, 梁瑞峰³, 杨亮¹

1. 河南医学高等专科学校,郑州 451191;
2. 郑州大学细胞生物研究室,郑州450008;
3. 河南省中医药研究院,郑州 450004

收稿日期:2016-08-25 出版日期:2017-03-26 发布日期:2017-03-07
作者简介:石科,女,博士,研究方向：生物工程;E-mail：shikesay@163.com,梁瑞峰为共同第一作者
基金资助:
河南省高等学校重点科研项目（16A310008）,河南省科技攻关计划项目（152102310161,162102310047）

Expression Analysis of Proteasome Subunit PSMD7 After Flagellar Disassembly of Dunaliella salina

SHI Ke^1,2, LIANG Rui-feng³, YANG Liang¹

1. Henan Medical College,Zhengzhou 451191;
2. Laboratory for Cell Biology of Zhengzhou University,Zhengzhou 450008;
3. Henan Province Chinese Medicine Research Institute,Zhengzhou 450004

Received:2016-08-25 Published:2017-03-26 Online:2017-03-07

摘要/Abstract

摘要： 为了研究盐藻19S蛋白酶体亚基PSMD7和鞭毛解聚的关系,用3-异丁基-1-甲基黄嘌呤（IBMX）诱导盐藻鞭毛解聚,实时荧光定量PCR检测PSMD7在鞭毛解聚后的mRNA表达变化。同时通过原核表达和蛋白纯化得到高纯度的盐藻PSMD7蛋白,制备盐藻PSMD7的山羊多克隆抗体,Western blot检测PSMD7蛋白在鞭毛解聚后的表达情况。结果显示IBMX诱导鞭毛解聚后PSMD7的mRNA表达水平增高,其中在鞭毛解聚后30 min达到最大值。成功制备了盐藻PSMD7的山羊多克隆抗体,Western blot结果显示PSMD7蛋白在鞭毛解聚后表达增加,说明蛋白酶体亚基PSMD7参与盐藻鞭毛的解聚。

关键词: PSMD7, 蛋白酶体, 鞭毛解聚

Abstract: To study the relationship between the 19S proteasome subunit PSMD7 and flagellar disassembly in Dunaliella salina,the flagellar disassembly was induced by 3-isobutyl-1-methylxanthine（IBMX）and the mRNA expression of PSMD7 was detected by real-time PCR. The high-purity PSMD7 protein of D. salina was obtained by prokaryotic expression and protein purification,followed by the preparation of goat anti-PSMD7 polyclonal antibody. The expression of PSMD7 protein after flagellar disassembly was investigated by Western blot. Results showed that the mRNA expression of PSMD7 increased after flagellar disassembly induced by IBMX and reached the highest at 30 min after disassembly. The goat anti-PSMD7 polyclonal antibody was successfully prepared. Results of Western blot showed that the protein level of PSMD7 rose after flagellar disassembly,indicating that the proteasome subunit PSMD7 was involved in flagellar disassembly of D. salina,thus this provides the basis for further study of the molecular mechanism of ubiquitin-proteasome system in flagellar disassembly.

Key words: PSMD7, proteasome, flagellar disassembly

石科, 梁瑞峰, 杨亮. 盐藻蛋白酶体亚基PSMD7在鞭毛解聚后的表达分析[J]. 生物技术通报, 2017, 33(3): 93-99.

SHI Ke, LIANG Rui-feng, YANG Liang. Expression Analysis of Proteasome Subunit PSMD7 After Flagellar Disassembly of Dunaliella salina[J]. Biotechnology Bulletin, 2017, 33(3): 93-99.

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盐藻蛋白酶体亚基PSMD7在鞭毛解聚后的表达分析

Expression Analysis of Proteasome Subunit PSMD7 After Flagellar Disassembly of Dunaliella salina

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