|本期目录/Table of Contents|

[1]韩芳,王志勇.日本囊对虾Ran基因的克隆表达与蛋白质GTP结合活性分析[J].集美大学学报(自然科学版),2010,15(4):241-247.
　HAN Fang,WANG Zhiyong.CloningRecombinant Expression and GTPbinding Activity Analysis of Ran Gene of Penaeus japonicus[J].Journal of Jimei University,2010,15(4):241-247.
点击复制

日本囊对虾Ran基因的克隆表达与蛋白质GTP结合活性分析(PDF)
分享到：

《集美大学学报（自然科学版）》[ISSN:1007-7405/CN:35-1186/N]

卷:
第15卷
期数:
2010年第4期
页码:
241-247
栏目:
水产科学与生物工程
出版日期:
2010-07-25

文章信息/Info

Title:
CloningRecombinant Expression and GTPbinding Activity Analysis of Ran Gene of Penaeus japonicus
作者:
韩芳1; 2; 王志勇1; 2
(1.集美大学水产学院,福建 厦门 361021;2.福建省高校水产科学技术与食品安全重点实验室,福建 厦门 361021)
Author(s):
HAN Fang1; 2; WANG Zhiyong1; 2
(1.Fisheries College,Jimei University,Xiamen 361021,China;2.Key Laboratory of Science and Technology forAquaculture and Food Safety(Jimei University),Fujian Province,Xiamen 361021,China)
关键词:
日本囊对虾; Ran基因; RACE﹣PCR; 蛋白表达; GTP活性
Keywords:
shrimp; Ran gene; RACEPCR; protein expression; GTP?binding activity
分类号:
-
DOI:
-
文献标志码:
-
摘要:
］在日本囊对虾抑制性差减杂交（suppression subtractive hybridization,SSH）研究过程中，首次发现一段经同源比较为Ran基因的部分序列，在抗病日本囊对虾中上调表达.为了进一步探索日本囊对虾Ran基因的功能，通过RACE-PCR的方法克隆得到了日本囊对虾Ran基因全长共1 441个碱基，其中开放阅读框为645个碱基，共编码215个氨基酸，这是首次在海洋无脊椎动物体内克隆到该基因.还将该基因克隆到原核表达载体PGEX-4T-2中并转化大肠杆菌BL21，37 ℃下诱导6 h，超声裂解表达菌株，结果表明GST-Ran融合蛋白在大肠杆菌中为可溶性表达，蛋白大小约为50 ku,经纯化得到了纯度大于90 %的GST-Ran融合蛋白.随后的GTP结合试验验证了Ran蛋白具有GTP结合活性
Abstract:
In previous studies,a cDNA fragment was found to be highly homologous with ras?related nuclear proteins(Ran proteins)by suppression subtractive hybridization(SSH)using WSSV?resistant shrimp.In this investigation，Ran gene was cloned from Penaeus japonicus shrimp by RACE-PCR，the total cDNA was 1 441 base pair and the ORF of shrimp Ran gene contained 645 bp，which encoded 215 amino acids．This was the first Ran gene obtained from marine invertebrates．Then the gene was cloned in pGEX-4T-2 and expressed in Escherichia coli BL21.After induction with IPTG at 37 ℃ for 6 h,the GST-Ran fusion protein(about 50 ku)was expressed and purified by glutathione?agarose beads.Based on protein domain analysis,the Ran protein was predicted to have a GTP/ATP binding motif.To test whether the Ran protein was endowed with GTPase activity,the GTP?binding assay was performed.The result revealed that the Ran protein had the GTP?binding activity

参考文献/References:

-

相似文献/References:

[1]岳亮,王艺磊,张子平,等.日本囊对虾性腺蛋白质双向电泳的样品制备方法的改进[J].集美大学学报(自然科学版),2011,16(4):246.
　YUE Liang,WANG Yi-lei,ZHANG Zi-ping,et al.A Novel Preparation Method of Marsupenaeus japonicus Gonad Samples for Two-dimensional Electrophoresis[J].Journal of Jimei University,2011,16(4):246.

备注/Memo

备注/Memo:
-
更新日期/Last Update: 2014-06-28