• 文献检索
  • 文档翻译
  • 深度研究
  • 学术资讯
  • Suppr Zotero 插件Zotero 插件
  • 邀请有礼
  • 套餐&价格
  • 历史记录
应用&插件
Suppr Zotero 插件Zotero 插件浏览器插件Mac 客户端Windows 客户端微信小程序
定价
高级版会员购买积分包购买API积分包
服务
文献检索文档翻译深度研究API 文档MCP 服务
关于我们
关于 Suppr公司介绍联系我们用户协议隐私条款
关注我们

Suppr 超能文献

核心技术专利：CN118964589B侵权必究
粤ICP备2023148730 号-1Suppr @ 2026

文献检索

告别复杂PubMed语法，用中文像聊天一样搜索，搜遍4000万医学文献。AI智能推荐，让科研检索更轻松。

立即免费搜索

文件翻译

保留排版，准确专业，支持PDF/Word/PPT等文件格式，支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述，25分钟生成高质量综述，智能提取关键信息，辅助科研写作。

立即免费体验

嗜热四膜虫的双功能二氢叶酸还原酶胸苷酸合成酶为分子和生物技术应用提供了一种工具。

The bifunctional dihydrofolate reductase thymidylate synthase of Tetrahymena thermophila provides a tool for molecular and biotechnology applications.

作者信息

Herrmann Lutz, Bockau Ulrike, Tiedtke Arno, Hartmann Marcus W W, Weide Thomas

机构信息

Cilian AG, Johann-Krane Weg 42, D-48149 Münster, Germany.

出版信息

BMC Biotechnol. 2006 Mar 20;6:21. doi: 10.1186/1472-6750-6-21.

DOI:10.1186/1472-6750-6-21
PMID:16549005
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1435751/
Abstract

BACKGROUND

Dihydrofolate reductase (DHFR) and thymidylate synthase (TS) are crucial enzymes in DNA synthesis. In alveolata both enzymes are expressed as one bifunctional enzyme.

RESULTS

Loss of this essential enzyme activities after successful allelic assortment of knock out alleles yields an auxotrophic marker in ciliates. Here the cloning, characterisation and functional analysis of Tetrahymena thermophila's DHFR-TS is presented. A first aspect of the presented work relates to destruction of DHFR-TS enzyme function in an alveolate thereby causing an auxotrophy for thymidine. A second aspect is to knock in an expression cassette encoding for a foreign gene with subsequent expression of the target protein.

CONCLUSION

This system avoids the use of antibiotics or other drugs and therefore is of high interest for biotechnological applications.

摘要

背景

二氢叶酸还原酶（DHFR）和胸苷酸合成酶（TS）是DNA合成中的关键酶。在肺泡虫中，这两种酶都作为一种双功能酶表达。

结果

在敲除等位基因成功进行等位基因分类后，这种必需酶活性的丧失在纤毛虫中产生了一个营养缺陷型标记。本文介绍了嗜热四膜虫DHFR-TS的克隆、表征和功能分析。所呈现工作的第一个方面涉及破坏肺泡虫中的DHFR-TS酶功能，从而导致对胸苷的营养缺陷。第二个方面是敲入一个编码外源基因的表达盒，随后表达目标蛋白。

结论

该系统避免了使用抗生素或其他药物，因此在生物技术应用中具有很高的价值。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9a05/1435751/7f6b9ef5e8b9/1472-6750-6-21-8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9a05/1435751/e4aa393b7c54/1472-6750-6-21-1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9a05/1435751/5c6afcf35640/1472-6750-6-21-2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9a05/1435751/3df96be0289c/1472-6750-6-21-3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9a05/1435751/9b2865a9a1f4/1472-6750-6-21-4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9a05/1435751/4d891c5ec337/1472-6750-6-21-5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9a05/1435751/f32691fd0936/1472-6750-6-21-6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9a05/1435751/959253cf3958/1472-6750-6-21-7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9a05/1435751/7f6b9ef5e8b9/1472-6750-6-21-8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9a05/1435751/e4aa393b7c54/1472-6750-6-21-1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9a05/1435751/5c6afcf35640/1472-6750-6-21-2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9a05/1435751/3df96be0289c/1472-6750-6-21-3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9a05/1435751/9b2865a9a1f4/1472-6750-6-21-4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9a05/1435751/4d891c5ec337/1472-6750-6-21-5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9a05/1435751/f32691fd0936/1472-6750-6-21-6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9a05/1435751/959253cf3958/1472-6750-6-21-7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9a05/1435751/7f6b9ef5e8b9/1472-6750-6-21-8.jpg

相似文献

1
The bifunctional dihydrofolate reductase thymidylate synthase of Tetrahymena thermophila provides a tool for molecular and biotechnology applications.嗜热四膜虫的双功能二氢叶酸还原酶胸苷酸合成酶为分子和生物技术应用提供了一种工具。
BMC Biotechnol. 2006 Mar 20;6:21. doi: 10.1186/1472-6750-6-21.
2
Primary structure of the dihydrofolate reductase-thymidylate synthase gene from Toxoplasma gondii.来自刚地弓形虫的二氢叶酸还原酶-胸苷酸合成酶基因的一级结构
J Biol Chem. 1993 Mar 25;268(9):6269-80.
3
Probing the role of parasite-specific, distant structural regions on communication and catalysis in the bifunctional thymidylate synthase-dihydrofolate reductase from Plasmodium falciparum.探究恶性疟原虫双功能胸苷酸合酶-二氢叶酸还原酶中寄生虫特异性的远距离结构区域在通讯和催化中的作用。
Biochemistry. 2008 Feb 5;47(5):1336-45. doi: 10.1021/bi701624u. Epub 2008 Jan 12.
4
Heterologous expression and characterization of the bifunctional dihydrofolate reductase-thymidylate synthase enzyme of Toxoplasma gondii.刚地弓形虫双功能二氢叶酸还原酶-胸苷酸合成酶的异源表达及特性分析
Biochemistry. 1996 May 21;35(20):6366-74. doi: 10.1021/bi952923q.
5
Human dihydrofolate reductase and thymidylate synthase form a complex in vitro and co-localize in normal and cancer cells.人二氢叶酸还原酶和胸苷酸合成酶在体外形成复合物，并在正常细胞和癌细胞中共定位。
J Biomol Struct Dyn. 2017 May;35(7):1474-1490. doi: 10.1080/07391102.2016.1186560. Epub 2016 Aug 5.
6
Kinetic reaction scheme for the dihydrofolate reductase domain of the bifunctional thymidylate synthase-dihydrofolate reductase from Leishmania major.来自硕大利什曼原虫的双功能胸苷酸合酶-二氢叶酸还原酶的二氢叶酸还原酶结构域的动力学反应方案。
Biochemistry. 1998 Sep 1;37(35):12206-12. doi: 10.1021/bi9803170.
7
First three-dimensional structure of Toxoplasma gondii thymidylate synthase-dihydrofolate reductase: insights for catalysis, interdomain interactions, and substrate channeling.刚地弓形虫胸苷酸合酶-二氢叶酸还原酶的首个三维结构：对催化、结构域间相互作用和底物通道的深入了解。
Biochemistry. 2013 Oct 15;52(41):7305-7317. doi: 10.1021/bi400576t. Epub 2013 Oct 3.
8
Disruption of the crossover helix impairs dihydrofolate reductase activity in the bifunctional enzyme TS-DHFR from Cryptosporidium hominis.交叉螺旋的破坏会损害来自人隐孢子虫的双功能酶TS-DHFR中的二氢叶酸还原酶活性。
Biochem J. 2009 Feb 1;417(3):757-64. doi: 10.1042/BJ20081247.
9
Functional expression of the dihydrofolate reductase domain of Leishmania major dihydrofolate reductase-thymidylate synthase bifunctional protein.硕大利什曼原虫二氢叶酸还原酶-胸苷酸合成酶双功能蛋白的二氢叶酸还原酶结构域的功能表达
Protein Expr Purif. 1996 Aug;8(1):23-7. doi: 10.1006/prep.1996.0070.
10
Cloning, nucleotide sequence and expression of the bifunctional dihydrofolate reductase-thymidylate synthase from Glycine max.大豆双功能二氢叶酸还原酶-胸苷酸合成酶的克隆、核苷酸序列及表达
Biochim Biophys Acta. 1995 Apr 26;1261(3):325-36. doi: 10.1016/0167-4781(94)00251-w.

引用本文的文献

1
Antibody production using a ciliate generates unusual antibody glycoforms displaying enhanced cell-killing activity.利用纤毛虫生产抗体可产生具有增强细胞杀伤活性的异常抗体糖型。
MAbs. 2016 Nov/Dec;8(8):1498-1511. doi: 10.1080/19420862.2016.1228504. Epub 2016 Sep 3.
2
Fusicoccins are biosynthesized by an unusual chimera diterpene synthase in fungi.壳梭孢菌素由真菌中一种不同寻常的嵌合二萜合酶生物合成。
Proc Natl Acad Sci U S A. 2007 Feb 27;104(9):3084-8. doi: 10.1073/pnas.0608426104. Epub 2007 Feb 20.
3
A recombinase system facilitates cloning of expression cassettes in the ciliate Tetrahymena thermophila.

本文引用的文献

1
Dynein: A Protein with Adenosine Triphosphatase Activity from Cilia.动力蛋白：纤毛中的一种具有三磷酸腺苷酶活性的蛋白质。
Science. 1965 Jul 23;149(3682):424-6. doi: 10.1126/science.149.3682.424.
2
Secretion of functional human enzymes by Tetrahymena thermophila.嗜热四膜虫分泌功能性人类酶。
BMC Biotechnol. 2006 Mar 16;6:19. doi: 10.1186/1472-6750-6-19.
3
RNA-guided DNA deletion in Tetrahymena: an RNAi-based mechanism for programmed genome rearrangements.四膜虫中RNA引导的DNA缺失：一种基于RNA干扰的程序性基因组重排机制。
一种重组酶系统有助于在嗜热四膜虫中克隆表达盒。
BMC Microbiol. 2007 Mar 1;7:12. doi: 10.1186/1471-2180-7-12.
Annu Rev Genet. 2005;39:537-59. doi: 10.1146/annurev.genet.39.073003.095906.
4
Tetrahymena thermophila.嗜热四膜虫
Curr Biol. 2005 May 10;15(9):R317-8. doi: 10.1016/j.cub.2005.04.039.
5
Programmed DNA deletion as an RNA-guided system of genome defense.作为一种RNA引导的基因组防御系统的程序性DNA缺失
Science. 2003 Jun 6;300(5625):1581-4. doi: 10.1126/science.1084737.
6
The circumsporozoite protein of Plasmodium falciparum is expressed and localized to the cell surface in the free-living ciliate Tetrahymena thermophila.恶性疟原虫的环子孢子蛋白在自由生活的嗜热四膜虫中表达并定位于细胞表面。
Mol Biochem Parasitol. 2002 Jul;122(2):119-26. doi: 10.1016/s0166-6851(02)00079-8.
7
Rooting the eukaryote tree by using a derived gene fusion.通过使用一个衍生的基因融合来确定真核生物树的根。
Science. 2002 Jul 5;297(5578):89-91. doi: 10.1126/science.1071196.
8
A robust inducible-repressible promoter greatly facilitates gene knockouts, conditional expression, and overexpression of homologous and heterologous genes in Tetrahymena thermophila.一个强大的可诱导-可抑制启动子极大地促进了嗜热四膜虫中基因敲除、条件表达以及同源和异源基因的过表达。
Proc Natl Acad Sci U S A. 2002 Mar 19;99(6):3734-9. doi: 10.1073/pnas.052016199. Epub 2002 Mar 12.
9
The I-antigens of Ichthyophthirius multifiliis are GPI-anchored proteins.多子小瓜虫的I抗原是糖基磷脂酰肌醇锚定蛋白。
J Eukaryot Microbiol. 2001 May-Jun;48(3):332-7. doi: 10.1111/j.1550-7408.2001.tb00322.x.
10
Knockout heterokaryons enable facile mutagenic analysis of essential genes in Tetrahymena.基因敲除异核体有助于对四膜虫中必需基因进行简便的诱变分析。
Methods Cell Biol. 2000;62:513-31. doi: 10.1016/s0091-679x(08)61554-x.