Difference between revisions of "Part:BBa K4162118"
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This biobrick was created through overlapping PCR of [https://parts.igem.org/Part:BBa_K4162009 BBa_K4162009](ribozyme+B0_RBS+crtE), [https://parts.igem.org/Part:BBa_K4162013 BBa_K4162013](ribozyme+T7_RBS+crtB), [https://parts.igem.org/Part:BBa_K4162016 BBa_K4162016](ribozyme+T7_RBS+crtI) and [https://parts.igem.org/Part:BBa_K4162019 BBa_K4162019](ribozyme+T7_RBS+crtY). These genes are a part of the carotenoid biosynthesis pathway and together, this biobrick converts farnesyl pyrophosphate to beta-carotene. In this part, the RNA sequences of hammerhead ribozyme conduct self-cleaving, and the polycistronic mRNA transcript is thus co-transcriptionally converted into individual mono-cistrons ''in vivo''. Self-interaction of the polycistron can be nullified and each cistron can initiate translation with comparable efficiency. | This biobrick was created through overlapping PCR of [https://parts.igem.org/Part:BBa_K4162009 BBa_K4162009](ribozyme+B0_RBS+crtE), [https://parts.igem.org/Part:BBa_K4162013 BBa_K4162013](ribozyme+T7_RBS+crtB), [https://parts.igem.org/Part:BBa_K4162016 BBa_K4162016](ribozyme+T7_RBS+crtI) and [https://parts.igem.org/Part:BBa_K4162019 BBa_K4162019](ribozyme+T7_RBS+crtY). These genes are a part of the carotenoid biosynthesis pathway and together, this biobrick converts farnesyl pyrophosphate to beta-carotene. In this part, the RNA sequences of hammerhead ribozyme conduct self-cleaving, and the polycistronic mRNA transcript is thus co-transcriptionally converted into individual mono-cistrons ''in vivo''. Self-interaction of the polycistron can be nullified and each cistron can initiate translation with comparable efficiency. | ||
+ | |||
+ | __TOC__ | ||
+ | |||
+ | ===Usage and Biology=== | ||
+ | |||
+ | We transfected this biobrick into ''E. coli'' to build single-cell factory for beta-carotene production. Coding sequences of crtEBIY are separated by ribozyme sequences. In this part, the RBS of crtBIY has equal intensity while the RBS of crtE is significantly stronger than the others. Since crtE catalyzes the first step of the carotenoid reaction chain, the concentration of substrate for the reaction catalyzed by this enzyme is significantly higher than for the next three steps of the reaction. To avoid more serious flux imbalance problems, we boosted the RBS intensity of crtE only in this biobrick and explored whether the carotenoid production of the strain could be significantly enhanced. | ||
+ | |||
+ | ===Characterization=== | ||
+ | |||
+ | ====定性1标题 ==== | ||
+ | |||
+ | 定性1文字 | ||
+ | |||
+ | [[File:T--Fudan--定性1图片不要用中文做文件名.png|400px|thumb|none|'''Figure 1. 图标题.''' 图注 引用的不要忘了写某某人某某年的出处]] | ||
+ | |||
+ | 定性1小结 | ||
+ | |||
+ | ====定性2标题 ==== | ||
+ | |||
+ | 定性2文字 | ||
+ | |||
+ | [[File:T--Fudan--定性2图片不要用中文做文件名.png|400px|thumb|none|'''Figure 2. 图标题.''' 图注 引用的不要忘了写某某人某某年的出处]] | ||
+ | |||
+ | 定性2小结 | ||
+ | |||
+ | |||
+ | ====定性3标题 ==== | ||
+ | |||
+ | 定性3文字 <ref>T7 phage factor required for managing RpoS in Escherichia coli. Tabib-Salazar A, Liu B, Barker D, Burchell L, Qimron U, Matthews SJ, Wigneshweraraj S. Proc Natl Acad Sci U S A, 2018 Jun 5;115(23):E5353-E5362. PMID:29789383</ref>. 又一个参考文献在一对尖括号内,例子里的参考文献格式是pubmed直接copy的 | ||
+ | |||
+ | [[File:T--Fudan--定性3图片不要用中文做文件名.png|400px|thumb|none|'''Figure 3. 图标题.''' 图注 引用的不要忘了写某某人某某年的出处]] | ||
+ | |||
+ | 定性3小结 | ||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here |
Revision as of 16:03, 11 October 2022
ribozyme+RBS+CDS module: crtEBIY
Introduction
This biobrick was created through overlapping PCR of BBa_K4162009(ribozyme+B0_RBS+crtE), BBa_K4162013(ribozyme+T7_RBS+crtB), BBa_K4162016(ribozyme+T7_RBS+crtI) and BBa_K4162019(ribozyme+T7_RBS+crtY). These genes are a part of the carotenoid biosynthesis pathway and together, this biobrick converts farnesyl pyrophosphate to beta-carotene. In this part, the RNA sequences of hammerhead ribozyme conduct self-cleaving, and the polycistronic mRNA transcript is thus co-transcriptionally converted into individual mono-cistrons in vivo. Self-interaction of the polycistron can be nullified and each cistron can initiate translation with comparable efficiency.
Usage and Biology
We transfected this biobrick into E. coli to build single-cell factory for beta-carotene production. Coding sequences of crtEBIY are separated by ribozyme sequences. In this part, the RBS of crtBIY has equal intensity while the RBS of crtE is significantly stronger than the others. Since crtE catalyzes the first step of the carotenoid reaction chain, the concentration of substrate for the reaction catalyzed by this enzyme is significantly higher than for the next three steps of the reaction. To avoid more serious flux imbalance problems, we boosted the RBS intensity of crtE only in this biobrick and explored whether the carotenoid production of the strain could be significantly enhanced.
Characterization
定性1标题
定性1文字
定性1小结
定性2标题
定性2文字
定性2小结
定性3标题
定性3文字 [1]. 又一个参考文献在一对尖括号内,例子里的参考文献格式是pubmed直接copy的
定性3小结
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 2202
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1681
Illegal NgoMIV site found at 1811
Illegal AgeI site found at 839 - 1000COMPATIBLE WITH RFC[1000]
- ↑ T7 phage factor required for managing RpoS in Escherichia coli. Tabib-Salazar A, Liu B, Barker D, Burchell L, Qimron U, Matthews SJ, Wigneshweraraj S. Proc Natl Acad Sci U S A, 2018 Jun 5;115(23):E5353-E5362. PMID:29789383