Sequence analysis and functional study of thymidylate synthase from zebrafish, Danio rerio
文献类型：期刊论文
| 作者 | Du, CQ; Niu, RL; Chu, E; Zhang, PJ; Lin, XK |
| 刊名 | JOURNAL OF BIOCHEMISTRY
 |
| 出版日期 | 2006-05-01 |
| 卷号 | 139期号:5页码:913-920 |
| 关键词 | Cloning And Expression Development Of Zebrafish Embryos Enzymatic Activity Thymidylate Synthase Zebrafish |
| ISSN号 | 0021-924X |
| DOI | 10.1093/jb/mvj100 |
| 文献子类 | Article |
| 英文摘要 | The thymidylate synthase (TS), an important target for many anticancer drugs, has been cloned from different species. But the cDNA property and function of TS in zebrafish are not well documented. In order to use zebrafish as an animal model for screening novel anticancer agents, we isolated TS cDNA from zebrafish and compared its sequence with those from other species. The open reading frame (ORF) of zebrafish TS cDNA sequence was 954 nucleotides, encoding a 318-amino acid protein with a calculated molecular mass of 36.15 kDa. The deduced amino acid sequence of zebrafish TS was similar to those from other organisms, including rat, mouse and humans. The zebrafish TS protein was expressed in Escherichia coli and purified to homogeneity. The purified zebrafish TS showed maximal activity at 28 degrees C with similar K-m value to human TS. Western immunoblot assay confirmed that TS was expressed in all the developmental stages of zebrafish with a high level of expression at the 1-4 cell stages. To study the function of TS in zebrafish embryo development, a short hairpin RNA (shRNA) expression vector, pSilencer 4.1-CMV/TS, was constructed which targeted the protein-coding region of zebrafish TS mRNA. Significant change in the development of tail and epiboly was found in zebrafish embryos microinjected pSilencer4.1-CMV/TS siRNA expression vector.; The thymidylate synthase (TS), an important target for many anticancer drugs, has been cloned from different species. But the cDNA property and function of TS in zebrafish are not well documented. In order to use zebrafish as an animal model for screening novel anticancer agents, we isolated TS cDNA from zebrafish and compared its sequence with those from other species. The open reading frame (ORF) of zebrafish TS cDNA sequence was 954 nucleotides, encoding a 318-amino acid protein with a calculated molecular mass of 36.15 kDa. The deduced amino acid sequence of zebrafish TS was similar to those from other organisms, including rat, mouse and humans. The zebrafish TS protein was expressed in Escherichia coli and purified to homogeneity. The purified zebrafish TS showed maximal activity at 28 degrees C with similar K-m value to human TS. Western immunoblot assay confirmed that TS was expressed in all the developmental stages of zebrafish with a high level of expression at the 1-4 cell stages. To study the function of TS in zebrafish embryo development, a short hairpin RNA (shRNA) expression vector, pSilencer 4.1-CMV/TS, was constructed which targeted the protein-coding region of zebrafish TS mRNA. Significant change in the development of tail and epiboly was found in zebrafish embryos microinjected pSilencer4.1-CMV/TS siRNA expression vector. |
| 学科主题 | Biochemistry & Molecular Biology |
| URL标识 | 查看原文 |
| 语种 | 英语 |
| WOS记录号 | WOS:000238357000012 |
| 公开日期 | 2010-12-24 |
| 源URL | [http://ir.qdio.ac.cn/handle/337002/5823]  |
| 专题 | 海洋研究所_实验海洋生物学重点实验室 |
| 作者单位 | 1.Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China 2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China 3.Yale Univ, Sch Med, Ctr Canc, New Haven, CT 06520 USA 4.Yale Univ, Sch Med, VACT Healthcare Syst, New Haven, CT 06520 USA |
| 推荐引用方式 GB/T 7714 | Du, CQ,Niu, RL,Chu, E,et al. Sequence analysis and functional study of thymidylate synthase from zebrafish, Danio rerio[J]. JOURNAL OF BIOCHEMISTRY,2006,139(5):913-920. |
| APA | Du, CQ,Niu, RL,Chu, E,Zhang, PJ,&Lin, XK.(2006).Sequence analysis and functional study of thymidylate synthase from zebrafish, Danio rerio.JOURNAL OF BIOCHEMISTRY,139(5),913-920. |
| MLA | Du, CQ,et al."Sequence analysis and functional study of thymidylate synthase from zebrafish, Danio rerio".JOURNAL OF BIOCHEMISTRY 139.5(2006):913-920. |
入库方式： OAI收割
来源：海洋研究所
浏览0
下载0
收藏0
其他版本
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。