Difference between revisions of "Os01g0777300"
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==Annotated Information== | ==Annotated Information== | ||
===Function=== | ===Function=== | ||
| − | + | In line #6, which showed approximately two blue spots per 100 mg of calli in the control, overexpression of I-SceI increased the number of GUS spots 2.3-fold, overexpression of OsExo1 increased GUS spots 3.6-fold and overexpression of OsRecQl4 increased GUS spots 2.5-fold. Overexpression of OsExo1or OsRecQl4 with I-SceI increased the number of GUS spots by 18- or 15-fold, respectively. Furthermore, we observed a synergistic effect of OsExo1 and OsRecQl4; a 29.8-fold increment was observed. However, in line #2, which showed approximately 20 blue spots per 100 mg of calli even in the control, no effects of the expression of each of the three genes were apparent. Nevertheless, both lines demonstrated that enhanced restoration of the GUS gene occurred upon DSB induction by I-SceI with OsRecQl4 or OsExo1. The results presented here suggest strongly that OsRecQl4 and OsExo1 enhance processing of the 50 end resection step after DSB induction by transiently expressed I-SceI and subsequent repair by HR and/or SSA in rice. The stable transformation of OsExo1 might be toxic for rice cells even when using the b-estradiol induction system. The overexpression of either OsRecQl4 or OsExo1 can enhance the processing of DSBs for efficient HR and SSA. BLM promotes resection of DNA strands in the 50-ssDNA resection step, and also removes Rad51 from ssDNA after invasion of the template. | |
===Expression=== | ===Expression=== | ||
| − | + | We investigated the frequency of GUS gene restoration using the following expression constructs: Pubi::OsExo1::TrbcS, Pubi::OsRecQl4::TrbcS and P2X35S::I-SceI::Thsp. The OsExo1 or OsRecQl4constructs were inserted into the pZK2 binary vector, OsExo1and I-SceI were inserted in the pZK2 binary vector, OsRecQl4and I-SceI were inserted in the pZD202 binary vector, and OsExo1, OsRecQl4and I-SceI were inserted into pZD202. | |
===Evolution=== | ===Evolution=== | ||
| − | |||
| − | + | Overexpression of OsRecQl4 and/or OsExo1 with the combination of I-SceI digestion further enhances the frequency of GUS restoration, probably due to enhanced resection of the I-SceI sites. The overexpression of either OsRecQl4 or OsExo1 alone, without I-SceI digestion, leads to increased numbers of GUS spots. This | |
| + | result suggests that spontaneously occurring DSBs in the GU–US construct were also progressively processed by overexpressed OsRecQl4 or OsExo1. | ||
==Labs working on this gene== | ==Labs working on this gene== | ||
Revision as of 14:00, 11 June 2014
Please input one-sentence summary here.
Contents
Annotated Information
Function
In line #6, which showed approximately two blue spots per 100 mg of calli in the control, overexpression of I-SceI increased the number of GUS spots 2.3-fold, overexpression of OsExo1 increased GUS spots 3.6-fold and overexpression of OsRecQl4 increased GUS spots 2.5-fold. Overexpression of OsExo1or OsRecQl4 with I-SceI increased the number of GUS spots by 18- or 15-fold, respectively. Furthermore, we observed a synergistic effect of OsExo1 and OsRecQl4; a 29.8-fold increment was observed. However, in line #2, which showed approximately 20 blue spots per 100 mg of calli even in the control, no effects of the expression of each of the three genes were apparent. Nevertheless, both lines demonstrated that enhanced restoration of the GUS gene occurred upon DSB induction by I-SceI with OsRecQl4 or OsExo1. The results presented here suggest strongly that OsRecQl4 and OsExo1 enhance processing of the 50 end resection step after DSB induction by transiently expressed I-SceI and subsequent repair by HR and/or SSA in rice. The stable transformation of OsExo1 might be toxic for rice cells even when using the b-estradiol induction system. The overexpression of either OsRecQl4 or OsExo1 can enhance the processing of DSBs for efficient HR and SSA. BLM promotes resection of DNA strands in the 50-ssDNA resection step, and also removes Rad51 from ssDNA after invasion of the template.
Expression
We investigated the frequency of GUS gene restoration using the following expression constructs: Pubi::OsExo1::TrbcS, Pubi::OsRecQl4::TrbcS and P2X35S::I-SceI::Thsp. The OsExo1 or OsRecQl4constructs were inserted into the pZK2 binary vector, OsExo1and I-SceI were inserted in the pZK2 binary vector, OsRecQl4and I-SceI were inserted in the pZD202 binary vector, and OsExo1, OsRecQl4and I-SceI were inserted into pZD202.
Evolution
Overexpression of OsRecQl4 and/or OsExo1 with the combination of I-SceI digestion further enhances the frequency of GUS restoration, probably due to enhanced resection of the I-SceI sites. The overexpression of either OsRecQl4 or OsExo1 alone, without I-SceI digestion, leads to increased numbers of GUS spots. This result suggests that spontaneously occurring DSBs in the GU–US construct were also progressively processed by overexpressed OsRecQl4 or OsExo1.
Labs working on this gene
Please input related labs here.
References
Please input cited references here.
Structured Information
| Gene Name |
Os01g0777300 |
|---|---|
| Description |
DNA repair protein (XPGC)/yeast Rad family protein |
| Version |
NM_001050956.2 GI:297597716 GeneID:4327899 |
| Length |
6382 bp |
| Definition |
Oryza sativa Japonica Group Os01g0777300, 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 1:34649248..34655629 |
| Sequence Coding Region |
34649281..34649441,34650177..34650296,34650566..34650689,34650812..34650949,34651080..34651289 |
| Expression | |
| Genome Context |
<gbrowseImage1> name=NC_008394:34649248..34655629 source=RiceChromosome01 preset=GeneLocation </gbrowseImage1> |
| Gene Structure |
<gbrowseImage2> name=NC_008394:34649248..34655629 source=RiceChromosome01 preset=GeneLocation </gbrowseImage2> |
| Coding Sequence |
<cdnaseq>atggggatccagggattgctgccgcagctgaagtcgatcatggcgcccatcggggtggaggcgctcaagggccagacggtggccgtcgacacctactcctggctccacaagggcgccctctcctgcggcgaccgcctctgcaagggcctacccaccaccaggcatattgagtattgcatgcatagggttaacatgttgcggcaccatggcgtgaaaccaatccttgtatttgacggaggtcatctgccgatgaagggtgatcaagagactaaacgtgaaaggtcacgaaaggaaaatcttgagcgtgccaaggagcatgaatctgctgggaattcacgtgctgcttttgagtgttatcagaaagctgttgacattacacctaggattgcttttgaactgatccaggtattgaagcaagaaaaggttgattatattgttgcgccttatgaagcagatgcacaaatgacattcttgtctgtgaacaaacttgttgatgccgttatcacagaggattctgatctgataccatttggctgttctagaattattttcaagatggacaagtttggacaaggtgttgaatttcatattacacgattgcagcgatgcagggagcttgatcttaatggattcactatgcaaatgctactagagatgtgtattttaagtggctgtgattatcttccatcattgcctggaatgggtgtgaaacgagctcatgcacttatccaaaagctcaagggtcatgaaaaggtaatcaagcacctgaggtacagtgctgtttctgttccacctcaatatgaagaaaacttcagaaaggcgatctgggcatttcagttccaaagagtttatgaccctgtgacggaagatattgttcatttgtcaggcattccccatggcagtagtgaagacttggactttttgggcccatggctaccacagactgttgctaaaggcattgcccaagggaatatcgatccgattaccaaggaaccatttgagggcaaaacagagagtagcgcactcgcttttgataaagtgcatctgaacagagagtccagtgctccttcaaatggaaagaaaaagcttgatctacctgtccaaagaaacgtattgacaaattacttctgcttagcatcgcttgaggcaaaacggaagttcagagcacctaaggttacacctaagcaacaagtacttaatgggtctttgcccagtcccagaattgaagactctggtacccctgatttaattgaagatactagcttgccatcaaataacattcaagtttatcagtgtagttctgagcattttagctcaggaacccctctagatgattcaatcaataccgcctctcaatgtagttctgagcgtgtccgctgtgacatccctcgggatgattcagccagtgtttcaccccagtgttctcatgatattggtagtgatcctgctgaggacccagacattgaaggcaacaaggtgaaagtcaacttctgcaacagaagtacaataccgacaggctctttcttagaggggactttgcctgggatatcagatccatttttagattcacacaacacagagccatccagagctgcaccacgttatgctgaaaaaagcaatgtggtctctgccaatagaaatattactgtgaggagttcatatttcaaaacagtaaataaaagagtttgcacaaatcaaggagaagatgagtgccatgatgaagataactgcgagactggtaattacactcttcctggagatcaacaaagaagctctggaggtatattgaagagaagaaaatttagtgatccccaaaattttgaagatggaatgtttcaacccactagtcctcatgaaagtccaccggttgctgataaaggctgtgactctgattcccacgacggtatcaacacaaattctgaaggaaaatttggatgcaatgtcgcccatgtaaataaatacagtggcattgcagagaaatcaatggataagtttgctgcgctgatatcatctttcagatacgcaggctctcgtgccagtggtcttcgcgctcctttaaaggatgtgaagaacactcttcctgtcagatctgttctcagacctccagaacagagattcggttgtacggctaagaagactactagagtgcctctacaaagcagattctcaagtgatgctacaaacagcactgacgttcctgatctgagcacatttgcatacaggcctacaactgcttctgctcattctgatcaggggaagatcacaagcaaagctacagatgctgctgctggacctcctgacctgagaacatttgcttatgcaccaacaagatctaccaccagccgttttgatcagagtgaaaacacgcgcaaagctatgtgcactgcggacagccctcctgatattagcacattcgaatataaacccatgaaaagcgccgtcagacgttcagatgggagcaagttttcaggtgcagcactaaaagcagctagacgaacctctcggagctga</cdnaseq> |
| Protein Sequence |
<aaseq>MGIQGLLPQLKSIMAPIGVEALKGQTVAVDTYSWLHKGALSCGD RLCKGLPTTRHIEYCMHRVNMLRHHGVKPILVFDGGHLPMKGDQETKRERSRKENLER AKEHESAGNSRAAFECYQKAVDITPRIAFELIQVLKQEKVDYIVAPYEADAQMTFLSV NKLVDAVITEDSDLIPFGCSRIIFKMDKFGQGVEFHITRLQRCRELDLNGFTMQMLLE MCILSGCDYLPSLPGMGVKRAHALIQKLKGHEKVIKHLRYSAVSVPPQYEENFRKAIW AFQFQRVYDPVTEDIVHLSGIPHGSSEDLDFLGPWLPQTVAKGIAQGNIDPITKEPFE GKTESSALAFDKVHLNRESSAPSNGKKKLDLPVQRNVLTNYFCLASLEAKRKFRAPKV TPKQQVLNGSLPSPRIEDSGTPDLIEDTSLPSNNIQVYQCSSEHFSSGTPLDDSINTA SQCSSERVRCDIPRDDSASVSPQCSHDIGSDPAEDPDIEGNKVKVNFCNRSTIPTGSF LEGTLPGISDPFLDSHNTEPSRAAPRYAEKSNVVSANRNITVRSSYFKTVNKRVCTNQ GEDECHDEDNCETGNYTLPGDQQRSSGGILKRRKFSDPQNFEDGMFQPTSPHESPPVA DKGCDSDSHDGINTNSEGKFGCNVAHVNKYSGIAEKSMDKFAALISSFRYAGSRASGL RAPLKDVKNTLPVRSVLRPPEQRFGCTAKKTTRVPLQSRFSSDATNSTDVPDLSTFAY RPTTASAHSDQGKITSKATDAAAGPPDLRTFAYAPTRSTTSRFDQSENTRKAMCTADS PPDISTFEYKPMKSAVRRSDGSKFSGAALKAARRTSRS</aaseq> |
| Gene Sequence |
<dnaseqindica>34..194#930..1049#1319..1442#1565..1702#1833..2042#2426..2600#2858..2931#3378..3504#3796..4142#4250..4304#4428..4723#4821..4872#5081..5288#5405..5828#gagagagagagagagagagagagagaggcggcaatggggatccagggattgctgccgcagctgaagtcgatcatggcgcccatcggggtggaggcgctcaagggccagacggtggccgtcgacacctactcctggctccacaagggcgccctctcctgcggcgaccgcctctgcaagggcctacccaccaccaggttcggctccggtgacctccccccttttcttggaacccagggtttcttaatccatttcgccgattggtgagacctaaatcgacggatccgggaaatctacttcttccccacgagcgtattgacagctgctggtcctaccaggggtttcttccgattttgttttcttggtcccgactggtgaaatccatttcggaggaggggtaaaaggcacgtcttgcttcggatgagagatttccgcattttttaggtgggtgtaggaaggggggggggatttcttgtttgttcagtctctctcaggattaaggtaataaatgtgcggcatttgttttcgcccccttccaggggcagtacatttactgaaatttgttatccgcctcagtgcccagattcttgtatgtctaggttgcattgaataaaatacacacttgggagagatccgctgcttggcatctggtgccgctttagttcagagaattcaccattttagtttgtggaatttttattggaaatagcttagacatgaatagttcaataaacgttaacaatagcttagcattgttagtagtatctttttactgtatgggatatcttctaacattgctgcatttttgatgtcgtgaactcataatgcggagggccatagatttgcaactccagtttttatcatcatttatcaccgttgtagttgagcaccataacttacaccttgtgtttcccctcctgaatctgaaccaggcatattgagtattgcatgcatagggttaacatgttgcggcaccatggcgtgaaaccaatccttgtatttgacggaggtcatctgccgatgaagggtgatcaagagactaaacgtgaaaggtatgttctttcttagggctccagaaagagcctaacatgtttcagttgccaaggaacatctacattacccttacagaacattaaaaaaaattgtgatttgacattttacatacggctatagttatggacaactgtggctttaggctgatatagagcagtcgcattattagtacagactagtggataagctaaaggcctccagccattctttgctatgtgatgtttcttgagtcattggatgctaagcaagtaagcatgacatcctgtaggtcacgaaaggaaaatcttgagcgtgccaaggagcatgaatctgctgggaattcacgtgctgcttttgagtgttatcagaaagctgttgacattacacctaggattgcttttgaactgatccaggtaaaggagcaaattccatgtatttttaccaaatatttaccacctatttgaagcagttctagtcaggagtaggaacatttggcttgtcttaactgattattgttccaaaacttgcaatgcaggtattgaagcaagaaaaggttgattatattgttgcgccttatgaagcagatgcacaaatgacattcttgtctgtgaacaaacttgttgatgccgttatcacagaggattctgatctgataccatttggctgttctagagtaagcacagtgttaaaacgctttatctttgtcctccacgtaccccatagtattttgctgctcagctagattagtgttcatagtttgttgtaacttctctaattaaaatgcatcttattccattgtgcagattattttcaagatggacaagtttggacaaggtgttgaatttcatattacacgattgcagcgatgcagggagcttgatcttaatggattcactatgcaaatgctactagagatgtgtattttaagtggctgtgattatcttccatcattgcctggaatgggtgtgaaacgagctcatgcacttatccaaaagctcaagggtcatgaaaaggtaaaccctgtatctgttgtatcaaatactaactttaggcctagttggaattggaaagaactgattcatgaactaacatgaacgatgcctgagatagtgttcaagacctacaaagcatggtaagaaaaggctggatagtagaaccagaatgcagttctacttagacataactacagaaggcaaggcctcaacacaaaattgattgcttcaacttttatgaggccaccgtcttgtacttaggtcattagccaatccatagcctgtcatacatgcaggacatgatttgagttactactgagcttgtagggcaacgatgtgcaatcactttcttgcttagcaaattagaaaaaaaaataatgagcaaatgatgcgtataatttcaggtaatcaagcacctgaggtacagtgctgtttctgttccacctcaatatgaagaaaacttcagaaaggcgatctgggcatttcagttccaaagagtttatgaccctgtgacggaagatattgttcatttgtcaggcattccccatggcagtagtgaagacttggactttttgggcccatatccttgccaaacttatatttgtacaatagatgctaaagctcattcatatgataacctgttgttttatccaccataagtccataaggatgaatttccacttttgtttggtcatgtctaatggccattatacaagttgcgttagaaagtaagctaacggcctttcaatgccaccatttctcagtaataccgaattgtgttacttgaagaaaatctcaacttgcatgcattggatcttttccttaattggtttatacatggctaccacagactgttgctaaaggcattgcccaagggaatatcgatccgattaccaaggaaccatttgaggtactaccgatcttccctcggttgttttggtttcttggtgctttgtttccttgaatgtaatttcatcctactctgattctgtgcactgcatatgtataaacatacccaagcatcaatatgcaaatacaaacgaggggctcttgaaaatggttgtaagctcactgaaatatccaatcagaggctacagagcattacagaagctcattagagaaaaacagataaaaatccataaaatattttgtaagaaaaaaatgtagggtcctcactgaaataagaaagtacattgtaagtgatcatggttttactgatgtactctgttttattaaagatcaccagtatgcaagcatttgctttgctgacatgcgaataccaacattcaattttatcttttgtggattgtcttactaagaagttatatatgttttatttgtatatgtgacttgcagggcaaaacagagagtagcgcactcgcttttgataaagtgcatctgaacagagagtccagtgctccttcaaatggaaagaaaaagcttgatctacctgtccaaagaaacgtattgacaaattacttctgtatcctctcaacacattttaatttaattgagagaacaatatatataccaatcaactttattatatgaactacttgttgcagcttatattatgggaatgccttggataagatagttcagttattgtgctttctcgttcccaaacatgtttaggatcaaatgaagaactaataatttccccataaacttatttacgaagtctaaggtgatgtgcgattcttagcatggtattttgcttgttttgtaaacaatacttcaactgtccattggatgaatccttgacttttagaaggcttagcatcgcttgaggcaaaacggaagttcagagcacctaaggttacacctaagcaacaagtacttaatgggtctttgcccagtcccagaattgaagactctggtacccctgatttaattgaagatactagcttgccatcaaataacattcaagtttatcagtgtagttctgagcattttagctcaggaacccctctagatgattcaatcaataccgcctctcaatgtagttctgagcgtgtccgctgtgacatccctcgggatgattcagccagtgtttcaccccagtgttctcatgatattggtagtgatcctgctgaggacccagacattgaaggcaacaaggtatttcacgtttctttattgccttattttcagtgttctcgtagtcctacctttctacaacaatatcgtatttccgtcattcttatagttttacttctgctatacaggtgaaagtcaacttctgcaacagaagtacaataccgacaggctctttcttagagggtaagagctctacctttttgtagttgatgcatgacttgatctgcaaatcatgtactggtatatctaattaaaatgaaaataatcttggtttctgttttgtatgcatttattttcgttttctagggactttgcctgggatatcagatccatttttagattcacacaacacagagccatccagagctgcaccacgttatgctgaaaaaagcaatgtggtctctgccaatagaaatattactgtgaggagttcatatttcaaaacagtaaataaaagagtttgcacaaatcaaggagaagatgagtgccatgatgaagataactgcgagactggtaattacactcttcctggagatcaacaaagaagctctggaggtatattgaagagaagaaaatttagtgatccccaaaattttgaagatgtaagcaatttttttttgagttgttattctatcatcatatcgcagtgaactgtctgtatcattctactatctggtcttgaatgtactatttgtgcagggaatgtttcaacccactagtcctcatgaaagtccaccggttgctgataaaggtaatagttccacctagttgtctgttcttattctctgtgagaatgtgcacctttccttgttccatttctctatagataaacttgtgacgattacaagaaattcagtttttccaaaaacaacagaaggttctgccaatgtttgagtgtgcgtagttccctttttttttgttgctgagggaaatcttctgaatgtctgatactaatgcaggctgtgactctgattcccacgacggtatcaacacaaattctgaaggaaaatttggatgcaatgtcgcccatgtaaataaatacagtggcattgcagagaaatcaatggataagtttgctgcgctgatatcatctttcagatacgcaggctctcgtgccagtggtcttcgcgctcctttaaaggatgtgaagaacactcttcctgtcaggtctgcagcttcagaaaatttcaattgcaccaagttcacacaatttacttatcagaacttaagagggggacacagttgtcccaatgagttgatatattttataacattaattgcagatctgttctcagacctccagaacagagattcggttgtacggctaagaagactactagagtgcctctacaaagcagattctcaagtgatgctacaaacagcactgacgttcctgatctgagcacatttgcatacaggcctacaactgcttctgctcattctgatcaggggaagatcacaagcaaagctacagatgctgctgctggacctcctgacctgagaacatttgcttatgcaccaacaagatctaccaccagccgttttgatcagagtgaaaacacgcgcaaagctatgtgcactgcggacagccctcctgatattagcacattcgaatataaacccatgaaaagcgccgtcagacgttcagatgggagcaagttttcaggtgcagcactaaaagcagctagacgaacctctcggagctgattctcacaaaattgtggatgcacctgccagcttccttatctgagcatgtttggatttgcgtatgccttctgtctgcgttcatgatctgagttcaacagaatcatccagcaacaaatgctctcaataaaagtgcattccattttgcccaaagtgcaagcacggctgaggtagctgccattcactactactagctgcagggcttatcagttcagacttcagagctcatccgtatctgcgcaatcagaagcacacatgttctgaagggaagtgctttatcttctcggtttccagagccaattatgtggcattttgcgagccaaatgtatgtgctcgtttttgccttgtccaatgaggttcttgtatgaacagcagtcgaatttgcgatgctatttaagccgcaaatgaggatactgttgtattgttcctttttgagaagatactgctgtatgttcattcttatacaagtgtgagcattccttcagaattcagatagcattactcttgtcaactattgatcaagatgttggaaagactttgaaaccggctgtttaagc</dnaseqindica> 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