Difference between revisions of "Os04g0661200"
(→Expression) |
(→Expression) |
||
| Line 7: | Line 7: | ||
===Expression=== | ===Expression=== | ||
| − | The transgenic plants generated were screened for the presence of the transgene cassette | + | The transgenic plants generated were screened for the presence of the transgene cassette by PCR analysis using wheat RGA2 intron-specific primer pairs (RGA2F, 5′-CCTGAAATTGGT AAAAGTAGA-3′, and RGA2R, 5′-TGTATCTTCATACTGCATTTG-3′). The genomic DNA from 21-day-old plants showed amplification of the wheat RGA2 intron only in the transgenic-positive plants, whereas no amplification was observed in the non-transgenic control (non-transformed Pusa Sugandhi II rice cultivar) plants. In the T0 generation, forty-five individual putative transgenic rice plants were generated of which approximately thirty were positive for the corresponding transgene, as confirmed by genomic PCR analysis. Among the positive transgenic plants screened, fourteen plants (T0) showing higher Pi levels were selected, and the T1 generation was produced . The transgenic plants (T1) exhibiting higher Pi levels (IO6–17, 82, 97, 112, and 163) were further selected, and subsequent generations (T2–T3) were grown under greenhouse conditions until maturity. After successive screening of the consequent generations (T1–T3), the progeny of IO6-97 (IO6-97-9-4) and IO6-163 (IO6-163-10-5) were selected. In the T3 generation, IO6-97-9-4 and IO6-163-10-5 showed a maximum Pi content (data not shown), thus all of the analysis were performed with the progeny of these two transgenic lines in the T4 generation. |
===Evolution=== | ===Evolution=== | ||
Revision as of 11:25, 4 June 2014
Please input one-sentence summary here.
Contents
Annotated Information
Function
The gene Os04g0661200 encodes myo-inositol-1,2,3,4,5,6-hexakisphosphate or IP6. Phytic acid (myo-inositol-1,2,3,4,5,6-hexakisphosphate or IP6) is known as the major source of phosphorus in cereal grains, comprising approximately 1–2% of the dry weight and accounting for approximately 65–80% of the total seed phosphorus.IP6 accumulates in the protein storage bodies as mixed salts called phytate that chelate a number of mineral cations. During the process of germination, endogenous grain phytase is activated, which degrades phytate, releasing stored phosphorus, myo-inositol and bound mineral cations that are further utilized by the developing seedlings. However, due to the lack of microbial phytase enzymes, monogastric animals are unable to remove the phosphates from the myo-inositol ring and are, therefore, incapable of utilizing the phosphorus present in cereals. Phytate has six negatively charged ions, making it a potent chelator of such divalent cations as Fe2+, Zn2+, Ca2+, and Mg2+ and rendering these ions unavailable for absorption by monogastric animals. In view of these adverse effects, many attempts have been made to reduce the phytic acid content in cereals.
Expression
The transgenic plants generated were screened for the presence of the transgene cassette by PCR analysis using wheat RGA2 intron-specific primer pairs (RGA2F, 5′-CCTGAAATTGGT AAAAGTAGA-3′, and RGA2R, 5′-TGTATCTTCATACTGCATTTG-3′). The genomic DNA from 21-day-old plants showed amplification of the wheat RGA2 intron only in the transgenic-positive plants, whereas no amplification was observed in the non-transgenic control (non-transformed Pusa Sugandhi II rice cultivar) plants. In the T0 generation, forty-five individual putative transgenic rice plants were generated of which approximately thirty were positive for the corresponding transgene, as confirmed by genomic PCR analysis. Among the positive transgenic plants screened, fourteen plants (T0) showing higher Pi levels were selected, and the T1 generation was produced . The transgenic plants (T1) exhibiting higher Pi levels (IO6–17, 82, 97, 112, and 163) were further selected, and subsequent generations (T2–T3) were grown under greenhouse conditions until maturity. After successive screening of the consequent generations (T1–T3), the progeny of IO6-97 (IO6-97-9-4) and IO6-163 (IO6-163-10-5) were selected. In the T3 generation, IO6-97-9-4 and IO6-163-10-5 showed a maximum Pi content (data not shown), thus all of the analysis were performed with the progeny of these two transgenic lines in the T4 generation.
Evolution
Please input evolution information here.
You can also add sub-section(s) at will.
Labs working on this gene
Please input related labs here.
References
Please input cited references here.
Structured Information
| Gene Name |
Os04g0661200 |
|---|---|
| Description |
Protein of unknown function DUF941 family protein |
| Version |
NM_001060682.1 GI:115461093 GeneID:4337286 |
| Length |
4234 bp |
| Definition |
Oryza sativa Japonica Group Os04g0661200, complete gene. |
| Source |
Oryza sativa Japonica Group ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
| Chromosome | |
| Location |
Chromosome 4:34133671..34137904 |
| Sequence Coding Region |
34134016..34134162,34134307..34134894,34135134..34135244,34135321..34135361,34135974..34136118 |
| Expression | |
| Genome Context |
<gbrowseImage1> name=NC_008397:34133671..34137904 source=RiceChromosome04 preset=GeneLocation </gbrowseImage1> |
| Gene Structure |
<gbrowseImage2> name=NC_008397:34133671..34137904 source=RiceChromosome04 preset=GeneLocation </gbrowseImage2> |
| Coding Sequence |
<cdnaseq>atggaggtggttctgcatgagggggatgccaaagattgggtttacaaaggagaaggtgcagcgaatctaatcctcagctacactggctcgtcaccttctatgcttggcaaggttctgcgagtcaaaaagattctaaaagacaagggacaaccggcaccaaactgtatagtcttctcaagtcatgaggaacatctgtggggcaagatcccaggattgttggaatctgttaaaaatgattgcttgccacaagcctatgctacaattgttatgagccaacatttgggtgccaatcatgttgatggcggggtccgtgtacgtgtgtctaagaactttttcgagcttgctggaaagaatgtgcttgacaaccgtcctgcatggagagtgaatgctagtgcaattgatgctggagctgattctgctcttctaatttctgaccacacattattttccggtaatcctagaggaagcagttgcatagcagtggagataaaggcaaaatgtggatttcttccgtcatctgaatacatatcgaaggagaattctatcaagaaacaagtaacacggtataagatgcatcagcacctaaaatttcatctgggagagatatcgaagacaagtgaatacgatcccctagatttattttctggatcaaaagaaagaatacatatggctatcaagtcatttttttcaactcctcagaacaactttagaatttttgtggatggttctttagtttttggtggcatgggaggtggcgcagatagtgttcatcctaatgaaacagagaaatgtcttgaagatctgagcaaggttactggcttacaactatctgactttattgagctcctgtcagaggcaatctttaagtctggagtgttgggtaaacttttagccactcaaaaactggatgatcatgacatcgaaggggcgattcatctgtactataacatcatttctcagccttgtttggtatgcaaaagtataactgatacagaacttctgcgcaagtactccaccttgcattctcttccattggacaagagtgagaagattgtcagggactttcttatctctgctactgcaaaagattgtagcctaatgatcagctttcgaccaagacagagtggaacaacagattctgagtacgattctgtatttcttgattcagtgaaccaaagctatgattacaaggcatattttattgatctggatgtgaaacccttggataagatggtacattactttaaattggatcaaaagatagtcaatttctacactagaaatggagaagttgggggagatccacgtgatcctccaaagggatgtggccctaggtga</cdnaseq> |
| Protein Sequence |
<aaseq>MEVVLHEGDAKDWVYKGEGAANLILSYTGSSPSMLGKVLRVKKI LKDKGQPAPNCIVFSSHEEHLWGKIPGLLESVKNDCLPQAYATIVMSQHLGANHVDGG VRVRVSKNFFELAGKNVLDNRPAWRVNASAIDAGADSALLISDHTLFSGNPRGSSCIA VEIKAKCGFLPSSEYISKENSIKKQVTRYKMHQHLKFHLGEISKTSEYDPLDLFSGSK ERIHMAIKSFFSTPQNNFRIFVDGSLVFGGMGGGADSVHPNETEKCLEDLSKVTGLQL SDFIELLSEAIFKSGVLGKLLATQKLDDHDIEGAIHLYYNIISQPCLVCKSITDTELL RKYSTLHSLPLDKSEKIVRDFLISATAKDCSLMISFRPRQSGTTDSEYDSVFLDSVNQ SYDYKAYFIDLDVKPLDKMVHYFKLDQKIVNFYTRNGEVGGDPRDPPKGCGPR</aaseq> |
| Gene Sequence |
<dnaseqindica>3743..3889#3011..3598#2661..2771#2544..2584#1787..1931#1388..1591#442..543#cgctttcctcatcgtcggctccacttccacttcctcttcctcctcctcgaggacgaggaggaaattggcagctggaggcttggagcgcgctgcatcatcccgcccgccctgctgctgcggcgatcgccggagcagggctgcaggggatccccacggccgcagcagctgttgttatctggtaagcaactaaagctcgctatctgctcgacggaatgccgcaccgaccaggaaccaagtgttgcttctttctcgcttgatcttctctttggtagatgagtaaatgaatgcacctgcacttggactcgagtttgctgtgttactcggcgttaagcagcaattttctgttcctcctacttcgtttttcgcctactattcctccttacttattttgaataatagaccttgaactcatacttggaagcctgcagattctgtgtggggatggaggtggttctgcatgagggggatgccaaagattgggtttacaaaggagaaggtgcagcgaatctaatcctcagctacactggctcgtcaccttctatggtatgcatcgaataggttctgcttgtgtcattattggttactgacctatagtctcccgtaaatcggcgaatttgatgcatgtgcttcaagaaggatgttttacttttgatgttctgatgattatagctatgaatgtttgggtgtctgggaggcgacagtagttcaagctcttttcatgagatgttttggatggtgttttatgaggtgatctggagatgtagccttgtgcttgatctagctaggacccagcagttggtgtctagtcgtttctgtcctagtttagctagctgtttttattttcctataagactctgtttgtttggctttcctcacaaagccggtttaatgaatgcctgatttccccccaaaaagatgttttaccaactttgatgcaacgcccagggatgcatgtcctatggccagagctgcactcttgaaatggcaaatgtttgtatatgtaaattagaaaatcccatacagctttcatgtagtcaaatagtttcactttaatattatataattcaaaagaaaataatcttagttgtgtgatcacacaaaaagaaggggtggtggcggcgccatcttagttgtgtgatctgcctgcccggtctagttttgtttttttgttcaattccttttgctgtattgctacaacattgttgtttcctgtaatggaaagagggagagcaaggttcttctgttttttaaaagaagagaggaaaaagagaaaataatcttatcttgacttggttgactaaaatttttagtttgtgaccaaagtattatgtttccctcgtatgatatgtttacaggaaaaatgttttgatgatgttttcttgaacagcttggcaaggttctgcgagtcaaaaagattctaaaagacaagggacaaccggcaccaaactgtatagtcttctcaagtcatgaggaacatctgtggggcaagatcccaggattgttggaatctgttaaaaatgattgcttgccacaagcctatgctacaattgttatgagccaacatttgggtgccaatcatgttgatggcggggtatgttttaaatccaattatttaatgttgttacggtgatatttattagaaataaatcaaccaacgttttccaatttaaaattcttttgctctataattaatggaaaattcgggttatttgaaaatacatccttgattagataattggatccacttgactaaaatgaataaacgtggtctctttttcatgttcaggtccgtgtacgtgtgtctaagaactttttcgagcttgctggaaagaatgtgcttgacaaccgtcctgcatggagagtgaatgctagtgcaattgatgctggagctgattctgctcttctaatttctgaccacacattattttccggtaaagtcttggcgtttcaatgttatttgaacgcttcatctctttcccttttgagtataatgtgactcttgtttgtactaagaaagagcaaatatcagttaggtctggtgatctattaactctttatgatataacatttgcagcaacactgttactcagtttattatttctatttgcatcacatgctttaaccttatttgtttgttaagagtggcaagtttgtaatttatggttttctagtctacaaccaaatagataggatcaagattggtaagcccaactttctccatgacacttgtgaaacttgggatatgtttggaaggattcatacaatttatatttctcaagattctccattattctgaaacatggaaagaatgaaagaacctgcttgttttttcgaatttctagcttaactctgatctgttccatccgcaagagcttgtttccgtcctaaaattggtagttggtccaaaattgattagtttgaatgattaaagatgattcggtagttttttacattgcgccattgctaaattattattcgtaactcgctgctagattttgttttttaacactatgtttgtttttggtaatttttctaaacctcaggtaatcctagaggaagcagttgcatagcagtggagataaaggtacattttgagctttatttatttcttcactgttcatcttttgtacacatttcttttaacatgctttgccttccaggcaaaatgtggatttcttccgtcatctgaatacatatcgaaggagaattctatcaagaaacaagtaacacggtataagatgcatcagcacctaaaatttcatctgggagaggtatgttggttttgggtaccagattttgatttcaccgcagccataattttgagtactaaatatataatagcacactactcttttgctcactgaaagacaacaaactactcttctagctgaaacagatattaatagatgcttcatattcattctgtcactcttggaagttgcaatgctatatcatcccttttgcttttcaaatcagcttggttctaaacatatttttaaacacatttcagatatcgaagacaagtgaatacgatcccctagatttattttctggatcaaaagaaagaatacatatggctatcaagtcatttttttcaactcctcagaacaactttagaatttttgtggatggttctttagtttttggtggcatgggaggtggcgcagatagtgttcatcctaatgaaacagagaaatgtcttgaagatctgagcaaggttactggcttacaactatctgactttattgagctcctgtcagaggcaatctttaagtctggagtgttgggtaaacttttagccactcaaaaactggatgatcatgacatcgaaggggcgattcatctgtactataacatcatttctcagccttgtttggtatgcaaaagtataactgatacagaacttctgcgcaagtactccaccttgcattctcttccattggacaagagtgagaagattgtcagggactttcttatctctgctactgcaaaagattgtagcctaatgatcagctttcgaccaagacagagtggaacaacagattctgagtacgattctgtatttcttgattcagtgaaccaaagctatgattacaaggtattgatgtgatatgtttcttgtttgagaagttcgataagtcatctagttccttgtggaacaattgtttttttctttctaaaaactctttttattaatgtttctttcccaccaacccattgatttgcacatttgttgttgcaggcatattttattgatctggatgtgaaacccttggataagatggtacattactttaaattggatcaaaagatagtcaatttctacactagaaatggagaagttgggggagatccacgtgatcctccaaagggatgtggccctaggtgacacaaaggttcagctccaaaggttcagctccaacattgacgaccttgaaatataaagggcataggacaagtgtctgttgtatcttggtgattgttgtgtttctacaagtgtgcatgtccaatgtgcaggttctcctcccttctagggaaaacgtgtaagagagtggtagttgttgtagtacatcctgtataccgtgtggtttgcctcagcagtaggccgaattttcagtagacaaacattaagaagagttaataaactgtatactgaggcatccacgtttaccttggaaatctatgataaattcccgtatactcagcttgctgtgaatctgtgatctgagttctg</dnaseqindica> |
| External Link(s) |
