Difference between revisions of "Part:BBa K1959000"
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<partinfo>BBa_K1959000 short</partinfo> | <partinfo>BBa_K1959000 short</partinfo> | ||
| − | The BBa_K1959000 harbors a coding sequence of phytoene synthase (PSY) derived from maize (''Zea mays''). Codon-optimization has been made for rice (''Oryza sativa''). PSY catalyzes the conversion of geranylgenranyl diphosphate to phytoene, which is a key enzyme for carotenoid biosynthesis. | + | The BBa_K1959000 harbors a coding sequence of phytoene synthase (PSY) derived from maize (''Zea mays''). Codon-optimization has been made for rice (''Oryza sativa''). PSY catalyzes the conversion of geranylgenranyl diphosphate (GGDP) to phytoene, which is a key enzyme for carotenoid biosynthesis. |
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===Usage and Biology=== | ===Usage and Biology=== | ||
| + | Phytoene synthase (PSY, EC 2.5.1.32) is an enzyme involved and responded for the first synthetic reaction in β-carotene biosynthetic pathway. In this reaction, two GGDP are converted to one molecule phytoene and two diphosphates (Figure. 1). ''PSY'' genes had been applied to the nutrition-improved crops such as Golden Rice. ''PSY'' gene of maize (''ZmPSY'') was introduced into rice genome to achieve the synthesis of β-carotene. In our project, the ''ZmPSY'' was used to reconstruct the pathway of astaxanthin biosynthesis. For maize is a monocots similar to rice, thus the ''ZmPSY'' was not fused with transit peptide for plastid sorting. | ||
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| + | ===Transcriptional activity of ''PSY''=== | ||
| + | Semi-quantitative RT-PCR was performed to detect the expression level of ''PSY'' involved in astaxanthin biosynthesis, total RNA of transgenic rice seeds were extracted and cDNA was synthesized from 1μg DNase-treated RNA. | ||
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Revision as of 17:15, 19 October 2016
Modification of Phytoene synthase (PSY)
The BBa_K1959000 harbors a coding sequence of phytoene synthase (PSY) derived from maize (Zea mays). Codon-optimization has been made for rice (Oryza sativa). PSY catalyzes the conversion of geranylgenranyl diphosphate (GGDP) to phytoene, which is a key enzyme for carotenoid biosynthesis.
Usage and Biology
Phytoene synthase (PSY, EC 2.5.1.32) is an enzyme involved and responded for the first synthetic reaction in β-carotene biosynthetic pathway. In this reaction, two GGDP are converted to one molecule phytoene and two diphosphates (Figure. 1). PSY genes had been applied to the nutrition-improved crops such as Golden Rice. PSY gene of maize (ZmPSY) was introduced into rice genome to achieve the synthesis of β-carotene. In our project, the ZmPSY was used to reconstruct the pathway of astaxanthin biosynthesis. For maize is a monocots similar to rice, thus the ZmPSY was not fused with transit peptide for plastid sorting.
Transcriptional activity of PSY
Semi-quantitative RT-PCR was performed to detect the expression level of PSY involved in astaxanthin biosynthesis, total RNA of transgenic rice seeds were extracted and cDNA was synthesized from 1μg DNase-treated RNA.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1045
Illegal NheI site found at 1164 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 105
Illegal AgeI site found at 956 - 1000COMPATIBLE WITH RFC[1000]