Difference between revisions of "Os03g0449200"
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| − | The OsAGO7 gene is the first isolated gene controlling upward curling of leaves in rice. | + | The OsAGO7 gene is the first isolated gene controlling upward curling of leaves in rice. It belongs to the AGO family because of its PAZ and PIWI domains. AGO is a large protein family that constitutes essential components of RNA-induced silencing complexes (RISCs), and their proteins are characterized by two unique domains, PAZ and PIWI (Carmell et al. 2002) |
In order to analyze the biological function of the OsAGO7 gene, Shi et al. joined a 223 bp fragment from the PAZ domain and a 510 bp fragment from the PIWI | In order to analyze the biological function of the OsAGO7 gene, Shi et al. joined a 223 bp fragment from the PAZ domain and a 510 bp fragment from the PIWI | ||
domain to the p1301UbiNos vector, in which there would be anti-sense segments expressed by the strong,Ubi promoter. The two resulting vectors—pAntiPAZ and pAntiPIWI—were transformed into the wild-type Zhonghua No. 11 by agrobacterium-mediated transformation.They obtained a total of 107 T0 transformants,with 80 being derived from pAntiPAZ and 27 from pAntiPIWI. In these transformants, we did not detect any obvious phenotype modiWcations in the T0 generation and the T1 generation. | domain to the p1301UbiNos vector, in which there would be anti-sense segments expressed by the strong,Ubi promoter. The two resulting vectors—pAntiPAZ and pAntiPIWI—were transformed into the wild-type Zhonghua No. 11 by agrobacterium-mediated transformation.They obtained a total of 107 T0 transformants,with 80 being derived from pAntiPAZ and 27 from pAntiPIWI. In these transformants, we did not detect any obvious phenotype modiWcations in the T0 generation and the T1 generation. | ||
| + | Since no phenotype changes in those anti-sense transformants were detected, we constructed an over-expression vector pCod, in which the whole OsAGO7 ORF was inserted into the 3� end of the Ubi promoter. After pCod was transformed into the wildtype Zhonghua No. 11, we found 58 T0 transformants, 52 (90%) of which exhibited the rolled-leaf trait. Furthermore, LRIs in the transformants were generally over 45 (Fig. 5a). Some exceeded 100, that is, the two margins of the leaves turned inside to overlap each other so that the whole leaf formed a cylinder (Fig. 5b). | ||
| + | To verify that the transformants were derived from | ||
| + | diVerent transformation events, we sampled several transformants with the rolled-leaf trait and isolated the Xanking sequences of inserted T-DNA left border by TAIL-PCR. This analysis conWrmed that T-DNA was inserted into diVerent sites in diVerent transformants. In order to further investigate if the insertion of T-DNA co-segregated with the rolled-leaf trait in the OsAGO7 over-expression transformants, we sampled three lines with the rolled-leaf trait and diVerent | ||
| + | T-DNA integration sites. Seeds were planted and raised to the T1 generation, and individuals were analyzed through GUS staining and PCR detection, where two pairs of primers—P3/Ubip and GusP+/GusP—were used (Fig. 3b). The results showed that the rolledleaf trait completely co-segregated with the integration | ||
| + | of T-DNA. | ||
===Expression=== | ===Expression=== | ||
Revision as of 06:14, 9 June 2014
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Contents
Annotated Information
Function
The OsAGO7 gene is the first isolated gene controlling upward curling of leaves in rice. It belongs to the AGO family because of its PAZ and PIWI domains. AGO is a large protein family that constitutes essential components of RNA-induced silencing complexes (RISCs), and their proteins are characterized by two unique domains, PAZ and PIWI (Carmell et al. 2002) In order to analyze the biological function of the OsAGO7 gene, Shi et al. joined a 223 bp fragment from the PAZ domain and a 510 bp fragment from the PIWI
domain to the p1301UbiNos vector, in which there would be anti-sense segments expressed by the strong,Ubi promoter. The two resulting vectors—pAntiPAZ and pAntiPIWI—were transformed into the wild-type Zhonghua No. 11 by agrobacterium-mediated transformation.They obtained a total of 107 T0 transformants,with 80 being derived from pAntiPAZ and 27 from pAntiPIWI. In these transformants, we did not detect any obvious phenotype modiWcations in the T0 generation and the T1 generation.
Since no phenotype changes in those anti-sense transformants were detected, we constructed an over-expression vector pCod, in which the whole OsAGO7 ORF was inserted into the 3� end of the Ubi promoter. After pCod was transformed into the wildtype Zhonghua No. 11, we found 58 T0 transformants, 52 (90%) of which exhibited the rolled-leaf trait. Furthermore, LRIs in the transformants were generally over 45 (Fig. 5a). Some exceeded 100, that is, the two margins of the leaves turned inside to overlap each other so that the whole leaf formed a cylinder (Fig. 5b). To verify that the transformants were derived from
diVerent transformation events, we sampled several transformants with the rolled-leaf trait and isolated the Xanking sequences of inserted T-DNA left border by TAIL-PCR. This analysis conWrmed that T-DNA was inserted into diVerent sites in diVerent transformants. In order to further investigate if the insertion of T-DNA co-segregated with the rolled-leaf trait in the OsAGO7 over-expression transformants, we sampled three lines with the rolled-leaf trait and diVerent T-DNA integration sites. Seeds were planted and raised to the T1 generation, and individuals were analyzed through GUS staining and PCR detection, where two pairs of primers—P3/Ubip and GusP+/GusP—were used (Fig. 3b). The results showed that the rolledleaf trait completely co-segregated with the integration of T-DNA.
Expression
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Evolution
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Labs working on this gene
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References
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Structured Information
| Gene Name |
Os03g0449200 |
|---|---|
| Description |
Stem cell self-renewal protein Piwi domain containing protein |
| Version |
NM_001057008.2 GI:297601171 GeneID:4333232 |
| Length |
4596 bp |
| Definition |
Oryza sativa Japonica Group Os03g0449200, 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 3:19879867..19884462 |
| Sequence Coding Region |
19882654..19884102 |
| Expression | |
| Genome Context |
<gbrowseImage1> name=NC_008396:19879867..19884462 source=RiceChromosome03 preset=GeneLocation </gbrowseImage1> |
| Gene Structure |
<gbrowseImage2> name=NC_008396:19879867..19884462 source=RiceChromosome03 preset=GeneLocation </gbrowseImage2> |
| Coding Sequence |
<cdnaseq>ctatcttttttttattgcagcgatacttatgctgatcagtttagtctccaagtgtcgaaacacatgacgaaactctctgggagggttctcttgcccccaaaattaaagcttggcagcagtgggcgcatcaaggacataacaccggaccgatttgatcgacagtggagctttctggatagccatgttgcagagggttccaagatcaagagttgggccttgataagttttggtggcaccccggagcagcacttttgcattacaaagtttgtgaaccagctatcaaatcggtgtgagcagctaggaattctgctaaacaagaagaccattatcagcccaatatttgagaggattcaactactcaataacgtgggaattttggagggcaagctcaagaaaattcaagaagctgcatcaggcaacttgcagttgctaatctgtgtcatggagaggaggcaccaaggctatgctgatctgaagcgaattgcagaaacatccattggtgttgtgacacaatgttgcctttactccaacttgagcaagctgacctctcaattcttgacgaatttggctttgaagatcaatgcgaaactcggtggctgcaatattgccctatacagcagctttccatgccaaattcctagaatatttttgtcggaggagccggtgatgttcatgggtgctgatgtcacacacccgcatccccttgatgattcaagtccatcagtggttgctgtagttgcaagcatgaattggccgtcagcaaataagtacatctccaggatgagatcacagacacaccggaaagaaatcattgagcaactggatgttatggctggtgaactgcttgaagagtttctaaaagaagtggggaagctcccaagcagaatcatattcttcagagatggtgtgagcgagacacagttctacaaggtgctgaaggaggagatgcatgcagtgcgcacaacttgttcgaggtatccgggttacaaacccttgatcacattcatcgtagttcagaagaggcatcacactagactcttccacagggagaggaatggcagctcgtcacactactctgatcagaacataccaccaggaacagttgtggacactgtgattacacacccaagggaatttgatttctatctgtgcagccactggggcaccaaggggacgagccggccaactcattatcatgttctgtgggatgagaataacttccgttccgacgaagtgcagcagttgatacacaatctttgctacacatttgctcggtgcaccaggccagtttctcttgtcccaccggcttactacgcacatctcgcggcatatagaggcaggctgtaccttgagaggtcagatacaactatgtacagggtcagtccgttgcagactgtgccgctacctaagctcagagacaatgttaagaggctcatgttctactgctag</cdnaseq> |
| Protein Sequence |
<aaseq>LSFFYCSDTYADQFSLQVSKHMTKLSGRVLLPPKLKLGSSGRIK DITPDRFDRQWSFLDSHVAEGSKIKSWALISFGGTPEQHFCITKFVNQLSNRCEQLGI LLNKKTIISPIFERIQLLNNVGILEGKLKKIQEAASGNLQLLICVMERRHQGYADLKR IAETSIGVVTQCCLYSNLSKLTSQFLTNLALKINAKLGGCNIALYSSFPCQIPRIFLS EEPVMFMGADVTHPHPLDDSSPSVVAVVASMNWPSANKYISRMRSQTHRKEIIEQLDV MAGELLEEFLKEVGKLPSRIIFFRDGVSETQFYKVLKEEMHAVRTTCSRYPGYKPLIT FIVVQKRHHTRLFHRERNGSSSHYSDQNIPPGTVVDTVITHPREFDFYLCSHWGTKGT SRPTHYHVLWDENNFRSDEVQQLIHNLCYTFARCTRPVSLVPPAYYAHLAAYRGRLYL ERSDTTMYRVSPLQTVPLPKLRDNVKRLMFYC</aaseq> |
| Gene Sequence |
<dnaseqindica>2788..4236#taccatccaaccatccaagaaccccctttctctctctctctctctctcaggcaagaaacggcgctcaagagcaagaagttgctgccatggcgtgttgatactcttactactaccaccaaaaagctgtgcacggacaggttaaagcggtgtaccgctgactcgcgcggccatggagggggagcgggagggggtggtggccaagaacgaggacaacgcaggcggaggtggtggtggattaggtacaggtggtaatggtggtggcggcggcggcggcagcgcgaacgggcggaggaggtggaggggcgggggcagcagcgggtataggcagcacccgatcatccaggcctacccggcgctcctgccgctgccgataaacggcgccaccggccacgcacacatcaacggcgcggtctcgctgccgctgcctctgccgccgccggtgctgctgtacctgcagccgccaccgccaccgccgctgctgcccctgctccccaaggtcgccgccgccacgttctacggcaagccacccaaggcggccgacgcggcaccgaggggctcaatgtggaagcacaggccatcgaaaaagccgccaccgcatgccattaccgccgcgctgctgccgctcccgcggggtaaggaaggcgctcctttttcttggttacttcccgccgcattgcgctgcggccacccatttaacacctcacaaagcagcgtgcaaacacaataactactagtactagttcatctctcctactagcatctactccatggtccatgctacactagctgctgttgcttttgttggattcttgatccatttacccttgccttgtagctcattttttattctcttcttccgattatgtctttatagggtgtgtttagttgctgaaaaaagttggaagtttaaagaaagtttggagtttggaaaaaaagttggaactaaacacgcccatatatatcaagaagtggttaatgtatagtagcaaacagttattataatggtaatagaaggatgatttgcaacagtattttgtgatcaatgcttcagtgccagcaagttttcttaggaagctgtggttttatggattcctgctgagattattatgtttcatcttgtgccgtcgctatcatactgtacttgcctgaggcagccaatgttgtgtgattattgatgataccttttcttttgaaagttagtaccccttcccctctcaaggaagggtctgaccatcttttaggaaggcactccctttttttaaccaatgatgtaccactatcttggtgtttgtgcatgccatagaaaatattattctacttattgatttattattgaatgatatttccctctaaccctgtcttttgatttataattttctctagatttaatcatcggtaaaactgtttaaatttcttcttcttctggctccttacatgaaggtattgatttgctaacagatggcaaggcgcttcaggagaaaatattcttcgcaaatgaaagaaaaacatctgaaaaggaggtaaatcatgtagacacccatgagaagtttactgttgcaccactggataatgcaatagcacggagacctgacatgggtggtgttgagggggccgagatccctctttctgcaaaccatttccttgtccagtttgatcctggccagaagattttccattacaatgttgacatatctccgcgtccatcaaaagaaacagcaagaatgatcaagaaaaaattagttgaagaaaatccaagtgttctctcaggttcccaaccagcctttgatggccgcaagaacttatatagtcctgttagatttcaggaggacagggttgagttctttgtcagcctcccagttgcattagcacgatgttcagtagttaaagaggatactggccacatgcttgacaaacagaaactcaagactttcaaagtgaatgtccggttggtttcgaagttatgtggtgaggacttgaacaaatatctgaacgaggacaaggatggcatccctcttccacaagattacctccatgcattggatgttgtgttgcgtgaaggtgctatggaaagttctatccttgtgggtcggtcactgtatgcacgctccatgggggaagcaagggacattggtggtggtgctgttggattaagaggtttctttcagagattgaggccaaccaagcaaggccttgcccttaatgttgatctctcactctcagctttccacgagagcacaggcataatttcatacttgcagaagcgctgtgacttcttgaaggaccttccacagaagaaaacaagggctttggcagaagaggagcacagggaggtggagaaagcattgaaaaatatccgggtatttgtgtgccatcgtgagactaatcaaaggtaccatgtgcatagcttgactaaggagacaacagagaacctcaagtttcgcgaccgaagtgggaaggatcttatggtggtggattacttcaaggagcactataaccatgatatacaattcaggaaccttccatgcttgcagattggcaggagcaagccatgttatgtgccaatggagctttgtgtagtttgtgagggccagaagtttcttggcaagctgtctgatgaacaaacttctaagattctgaaaatgggttgtgaaagaccgagtgaaagaaagggaatcataaagggtgttgtcaaaggtgcatttcatgcaagaaggtatggttcttcattcttgtcattttcttttgcagcagttttgagctatagaaatcacataaatgtaagcgtagctatcttttttttattgcagcgatacttatgctgatcagtttagtctccaagtgtcgaaacacatgacgaaactctctgggagggttctcttgcccccaaaattaaagcttggcagcagtgggcgcatcaaggacataacaccggaccgatttgatcgacagtggagctttctggatagccatgttgcagagggttccaagatcaagagttgggccttgataagttttggtggcaccccggagcagcacttttgcattacaaagtttgtgaaccagctatcaaatcggtgtgagcagctaggaattctgctaaacaagaagaccattatcagcccaatatttgagaggattcaactactcaataacgtgggaattttggagggcaagctcaagaaaattcaagaagctgcatcaggcaacttgcagttgctaatctgtgtcatggagaggaggcaccaaggctatgctgatctgaagcgaattgcagaaacatccattggtgttgtgacacaatgttgcctttactccaacttgagcaagctgacctctcaattcttgacgaatttggctttgaagatcaatgcgaaactcggtggctgcaatattgccctatacagcagctttccatgccaaattcctagaatatttttgtcggaggagccggtgatgttcatgggtgctgatgtcacacacccgcatccccttgatgattcaagtccatcagtggttgctgtagttgcaagcatgaattggccgtcagcaaataagtacatctccaggatgagatcacagacacaccggaaagaaatcattgagcaactggatgttatggctggtgaactgcttgaagagtttctaaaagaagtggggaagctcccaagcagaatcatattcttcagagatggtgtgagcgagacacagttctacaaggtgctgaaggaggagatgcatgcagtgcgcacaacttgttcgaggtatccgggttacaaacccttgatcacattcatcgtagttcagaagaggcatcacactagactcttccacagggagaggaatggcagctcgtcacactactctgatcagaacataccaccaggaacagttgtggacactgtgattacacacccaagggaatttgatttctatctgtgcagccactggggcaccaaggggacgagccggccaactcattatcatgttctgtgggatgagaataacttccgttccgacgaagtgcagcagttgatacacaatctttgctacacatttgctcggtgcaccaggccagtttctcttgtcccaccggcttactacgcacatctcgcggcatatagaggcaggctgtaccttgagaggtcagatacaactatgtacagggtcagtccgttgcagactgtgccgctacctaagctcagagacaatgttaagaggctcatgttctactgctagcttcaccgccctgaatttcatggtatcgggactctcctacagtaattttcctcccaaaaaaaaaggtgtttcagatgttatctttaacgtatatcgcctgtcatcgtgtttttctatatttgtccgaattttggaaaatatgtatagtgctcctggacagaaatcctgtgcatcataacatcttacagcagttggcacttagtatttatgcaaaaattctgaaaccaaactgtttcattcccatagatgatgttggtaaggtgggggtttacctatttgtacagtctggatagtgaggaggtcctggagatgaagatactgctgtcgtgttccaaatttaatttcgtttaaattactgct</dnaseqindica> |
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