Difference between revisions of "Os03g0230500"
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| − | + | The rice gene ''Os03g0230500'' was reported as '''''DSM3''''' in 2011<ref name="ref1"/>, '''''OsITPK2'''''('''''OsITP5/6K-2''''') in 2007<ref name="ref2"/>, respectively. | |
==Annotated Information== | ==Annotated Information== | ||
===Function=== | ===Function=== | ||
| − | + | *''DSM3'' is predicted to '''encode a putative ITPK''' with 349 amino acids and a predicted molecular mass of 38.8 kDa. ''DSM3''/ ''OsITPK2'' is an important '''member''' of the '''OsITPK family '''for stress responses, and an optimal expression level is '''essential for drought''' and '''salt tolerance''' in rice<ref name="ref1"/>. | |
| + | |||
| + | *According to the predicted function of ''DSM3'' as '''an ITPK''' and the '''downstream genes affected''', ''DSM3'' may contribute significantly in '''inositol phosphate–mediated stress signal transductions''', although its biochemical characteristics and transcriptional regulation related to other physiological substrates and products remain to be identified<ref name="ref1"/>. | ||
| + | |||
| + | *Disruption or overexpression of ''DSM3'' can '''affect''' the expression of some '''stress-responsive genes''' and some of the '''homologous genes'''<ref name="ref1"/>: | ||
| + | **Two '''peroxidase genes''' (''OsPOX8.1'' and ''OsPOX22.3'') showed slightly higher expression levels in both the mutant and overexpression lines than in the WT' before drought stress, but their drought-induced expression levels were '''lower''' than that in the WT'. | ||
| + | **''Du et al.'' checked a few '''osmotic adjustment–related genes''' (''OsP5CS'', ''OsLEA3'', ''OsRAB16b'', and ''OsGDSL'') and the results showed that the ''OsP5CS'' and ''OsGDSL'' had significantly '''lower levels''' in both the mutant and overexpression lines under drought stress conditions. | ||
| + | |||
| + | ===Mutation=== | ||
| + | *a '''drought'''- and '''salt'''-'''hypersensitive''' mutant ''dsm3''<ref name="ref1"/>: | ||
| + | **The mutant phenotype was caused by a '''T-DNA insertion''' in a gene encoding a putative inositol 1,3,4-trisphosphate 5/6-kinase previously named ''OsITPK2'' with unknown function. | ||
| + | **Under drought stress conditions, the mutant had significantly less accumulation of osmolytes such as proline and soluble sugar and showed significantly reduced root volume, spikelet fertility, biomass, and grain yield; however, malondialdehyde level was increased in the mutant. | ||
| + | **A few genes related to osmotic adjustment and reactive oxygen species scavenging were down-regulated in the mutant and overexpression lines. | ||
| + | *positive and negative lines<ref name="ref1"/>: | ||
| + | **Three positive (DSM3-suppressed) artificial miRNA lines | ||
| + | ***ai-1 | ||
| + | ***ai-9 | ||
| + | ***ai-14 | ||
| + | **three negative lines | ||
| + | ***ai-3 | ||
| + | ***ai-4 | ||
| + | ***ai-13 | ||
| + | **They were selected for drought and salt stress testing. The results showed that the positive lines were '''hypersensitive''' to drought and salt stresses , which is in agreement with the phenotype of the ''dsm3'' mutant. | ||
| + | **The '''survival rate''' of the positive amiR-''DSM3'' lines was only 5–15%, significantly '''lower''' than that of the control (50–70%). | ||
| + | **In the '''salt treatment''', the '''plant height''' of the positive amiR-DSM3 lines was significantly lower than that of the control. | ||
===Expression=== | ===Expression=== | ||
| − | + | *Overexpression of ''DSM3'' (''OsITPK2'') in rice resulted in '''drought- and salt-hypersensitive phenotypes''' and '''physiological changes''' similar to those in the mutant. '''Inositol trisphosphate''' (IP3) '''level''' was '''decreased''' in the overexpressors under normal condition and drought stress<ref name="ref1"/>. | |
| + | |||
| + | *The expression level of ''DSM3'' promoter-driven b-glucuronidase (GUS) reporter gene in rice was '''induced by drought''', '''salt''' and '''abscisic acid'''. Transcript level analysis of OsITPK genes revealed that they had '''different tempo-spatial expression patterns''', and the responses of ''DSM3'' to abiotic stresses, including drought, salinity, cold, and high temperature, were distinct from the other five members in rice<ref name="ref1"/>. | ||
| + | |||
| + | ===Subcellular localization=== | ||
| + | '''ER localization''' of ''DSM3'' was confirmed by transient expression in ''Arabidopsis'' mesophyll | ||
| + | protoplasts. The result of ER localization of the ''DSM3'' was consistent with its biological process as a '''functional ITPK''' in rice<ref name="ref1"/>. | ||
===Evolution=== | ===Evolution=== | ||
| − | + | [[File: OsITPK Phylogenetic1.jpg|right|thumb|250px|'''Figure 1.''' ''Phylogenetic analysis of ITPK families and exon–intron structures of OsITPK genes.(from reference <ref name="ref1"/>).'']] | |
| − | + | Sequence analysis revealed '''six putative ITPKs in rice'''<ref name="ref1"/><ref name="ref2"/>. | |
| − | + | *Phylogenetic analysis<ref name="ref1"/>: | |
| + | **Phylogenetic analysis based on the deduced protein sequences revealed '''three subgroups''' for these ITPKs (Fig. 1a). | ||
| + | ***OsITPKs are distributed in all the three subgroups. ''OsITPK1'', ''OsITPK2'', and ''OsITPK3'' were clustered in '''subgroup I'''. | ||
| + | ***''OsITPK4'' and ''OsITPK5'' were classified into '''subgroup II''. | ||
| + | ***''OsITPK6'' resided in '''subgroup III''', in which ITPKs from vertebrates were predominant (Fig. 1a). | ||
| + | **The ITPKs from ''Arabidopsis'' were also distributed in all three subgroups. | ||
| + | *the exon–intron structures of the OsITPK genes and their chromosomal locations<ref name="ref1"/>: | ||
| + | **''OsITPK4'' and ''OsITPK5'' belonging to '''subgroup II''' (Fig. 1A), have no intron (Fig. 1b). | ||
| + | **''OsITPK6'' contained 12 exons and 11 introns and belongs to '''subgroup III'''. | ||
| + | **The other three rice ITPKs (''OsITPK1'' to ''OsITPK3'') belong to '''subgroup I''', with each containing 10 exons and 9 introns with very similar intron phases (Fig. 1b). | ||
| + | **The exon–intron organization structures of the ''OsITPK'' genes suggest that ''OsITPK1'', ''OsITPK2'', and ''OsITPK3'' are '''similar to each other''' (Fig. 1b). | ||
==Labs working on this gene== | ==Labs working on this gene== | ||
| − | + | *National Key Laboratory of Crop Genetic Improvement and National Center of Plant Gene Research (Wuhan), Huazhong Agricultural University, 430070 Wuhan, China | |
==References== | ==References== | ||
| − | + | <references> | |
| + | * <ref name="ref1"> | ||
| + | Du H, Liu L, You L, et al. Characterization of an inositol 1, 3, 4-trisphosphate 5/6-kinase gene that is essential for drought and salt stress responses in rice[J]. Plant molecular biology, 2011, 77(6): 547-563. | ||
| + | </ref> | ||
| + | * <ref name="ref2"> | ||
| + | Suzuki M, Tanaka K, Kuwano M, et al. Expression pattern of inositol phosphate-related enzymes in rice (Oryza sativa L.): implications for the phytic acid biosynthetic pathway[J]. Gene, 2007, 405(1): 55-64. | ||
| + | </references> | ||
| + | |||
==Structured Information== | ==Structured Information== | ||
Revision as of 08:43, 3 March 2015
The rice gene Os03g0230500 was reported as DSM3 in 2011[1], OsITPK2(OsITP5/6K-2) in 2007[2], respectively.
Contents
Annotated Information
Function
- DSM3 is predicted to encode a putative ITPK with 349 amino acids and a predicted molecular mass of 38.8 kDa. DSM3/ OsITPK2 is an important member of the OsITPK family for stress responses, and an optimal expression level is essential for drought and salt tolerance in rice[1].
- According to the predicted function of DSM3 as an ITPK and the downstream genes affected, DSM3 may contribute significantly in inositol phosphate–mediated stress signal transductions, although its biochemical characteristics and transcriptional regulation related to other physiological substrates and products remain to be identified[1].
- Disruption or overexpression of DSM3 can affect the expression of some stress-responsive genes and some of the homologous genes[1]:
- Two peroxidase genes (OsPOX8.1 and OsPOX22.3) showed slightly higher expression levels in both the mutant and overexpression lines than in the WT' before drought stress, but their drought-induced expression levels were lower than that in the WT'.
- Du et al. checked a few osmotic adjustment–related genes (OsP5CS, OsLEA3, OsRAB16b, and OsGDSL) and the results showed that the OsP5CS and OsGDSL had significantly lower levels in both the mutant and overexpression lines under drought stress conditions.
Mutation
- a drought- and salt-hypersensitive mutant dsm3[1]:
- The mutant phenotype was caused by a T-DNA insertion in a gene encoding a putative inositol 1,3,4-trisphosphate 5/6-kinase previously named OsITPK2 with unknown function.
- Under drought stress conditions, the mutant had significantly less accumulation of osmolytes such as proline and soluble sugar and showed significantly reduced root volume, spikelet fertility, biomass, and grain yield; however, malondialdehyde level was increased in the mutant.
- A few genes related to osmotic adjustment and reactive oxygen species scavenging were down-regulated in the mutant and overexpression lines.
- positive and negative lines[1]:
- Three positive (DSM3-suppressed) artificial miRNA lines
- ai-1
- ai-9
- ai-14
- three negative lines
- ai-3
- ai-4
- ai-13
- They were selected for drought and salt stress testing. The results showed that the positive lines were hypersensitive to drought and salt stresses , which is in agreement with the phenotype of the dsm3 mutant.
- The survival rate of the positive amiR-DSM3 lines was only 5–15%, significantly lower than that of the control (50–70%).
- In the salt treatment, the plant height of the positive amiR-DSM3 lines was significantly lower than that of the control.
- Three positive (DSM3-suppressed) artificial miRNA lines
Expression
- Overexpression of DSM3 (OsITPK2) in rice resulted in drought- and salt-hypersensitive phenotypes and physiological changes similar to those in the mutant. Inositol trisphosphate (IP3) level was decreased in the overexpressors under normal condition and drought stress[1].
- The expression level of DSM3 promoter-driven b-glucuronidase (GUS) reporter gene in rice was induced by drought, salt and abscisic acid. Transcript level analysis of OsITPK genes revealed that they had different tempo-spatial expression patterns, and the responses of DSM3 to abiotic stresses, including drought, salinity, cold, and high temperature, were distinct from the other five members in rice[1].
Subcellular localization
ER localization of DSM3 was confirmed by transient expression in Arabidopsis mesophyll protoplasts. The result of ER localization of the DSM3 was consistent with its biological process as a functional ITPK in rice[1].
Evolution
Sequence analysis revealed six putative ITPKs in rice[1][2].
- Phylogenetic analysis[1]:
- Phylogenetic analysis based on the deduced protein sequences revealed three subgroups for these ITPKs (Fig. 1a).
- OsITPKs are distributed in all the three subgroups. OsITPK1, OsITPK2, and OsITPK3 were clustered in subgroup I.
- OsITPK4 and OsITPK5 were classified into 'subgroup II.
- OsITPK6 resided in subgroup III, in which ITPKs from vertebrates were predominant (Fig. 1a).
- The ITPKs from Arabidopsis were also distributed in all three subgroups.
- Phylogenetic analysis based on the deduced protein sequences revealed three subgroups for these ITPKs (Fig. 1a).
- the exon–intron structures of the OsITPK genes and their chromosomal locations[1]:
- OsITPK4 and OsITPK5 belonging to subgroup II (Fig. 1A), have no intron (Fig. 1b).
- OsITPK6 contained 12 exons and 11 introns and belongs to subgroup III.
- The other three rice ITPKs (OsITPK1 to OsITPK3) belong to subgroup I, with each containing 10 exons and 9 introns with very similar intron phases (Fig. 1b).
- The exon–intron organization structures of the OsITPK genes suggest that OsITPK1, OsITPK2, and OsITPK3 are similar to each other (Fig. 1b).
Labs working on this gene
- National Key Laboratory of Crop Genetic Improvement and National Center of Plant Gene Research (Wuhan), Huazhong Agricultural University, 430070 Wuhan, China
References
- ↑ 1.00 1.01 1.02 1.03 1.04 1.05 1.06 1.07 1.08 1.09 1.10 1.11 1.12 Du H, Liu L, You L, et al. Characterization of an inositol 1, 3, 4-trisphosphate 5/6-kinase gene that is essential for drought and salt stress responses in rice[J]. Plant molecular biology, 2011, 77(6): 547-563.
- ↑ 2.0 2.1 Cite error: Invalid
<ref>tag; no text was provided for refs namedref2
Structured Information
| Gene Name |
Os03g0230500 |
|---|---|
| Description |
Inositol 1, 3, 4-trisphosphate 56-kinase family protein |
| Version |
NM_001055992.1 GI:115451712 GeneID:4332146 |
| Length |
5292 bp |
| Definition |
Oryza sativa Japonica Group Os03g0230500, 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 3:6951443..6956734 |
| Sequence Coding Region |
6951685..6951872,6953287..6953368,6953555..6953587,6953887..6954007,6954778..6954870 |
| Expression | |
| Genome Context |
<gbrowseImage1> name=NC_008396:6951443..6956734 source=RiceChromosome03 preset=GeneLocation </gbrowseImage1> |
| Gene Structure |
<gbrowseImage2> name=NC_008396:6951443..6956734 source=RiceChromosome03 preset=GeneLocation </gbrowseImage2> |
| Coding Sequence |
<cdnaseq>atgcggctgcacggggaggtttccttcgatgaggacgaggaggaagtggtgatggttccggcggcggcgctgtcgtcgtcgccgctcaacggcggggccgttccggtgacgaggctcgtggtggggtacgccctcaccaagaagaaggtgaagagcttccttcagcccaatctgctgttgctggcgaggaagaagggaattaatcttgtagcaattgatgacactcgcccgcttgcagaacaaggcccatttgatgttatcttgcacaagattactagcaaggaatggcaacaggttctggaggattatcatgaggaacatccagaggttactgtccttgacccaccaaatgccatcaatcatctgaataatcggcaatctatgcttgcagaagtatctgatttgaacttatccagtttctatggagaagtttgcactccacgccaactggtcattatgagagatccatcctctataccaaccgcagttgccatggctggactaaccttgcccttggttgcgaagccattggttgttgatggaacatctaaatcccacgaactatctcttgcatatgatgaggcatccttatcaatgcttgatcctcctctggtcctccaggaatttgtgaaccatggtgggatcctctttaaggtgtacatcattggtgaaactatacaagttgtacggaggttttctcttcctgatgttaacacttatgacttattaaacaatgttggcgtctatcgatttccaagagtttcatgtgctgcagctagtgcagaccatgcagacctcgatcctcatatctcagaacttcctccaagaccactcctagagaaactgggaaaagagcttcgtggaagactgggtttaagattgttcaacatagatatgatcagagaacttggaaccaaagatcggtactatataattgacatcaactacttcccagggttcgggaaaatgccaggttatgagcacatattcaccgatttcttgctgaatcttgcgcaaagcaagtacaagaagtgcttaagcggcggctga</cdnaseq> |
| Protein Sequence |
<aaseq>MRLHGEVSFDEDEEEVVMVPAAALSSSPLNGGAVPVTRLVVGYA LTKKKVKSFLQPNLLLLARKKGINLVAIDDTRPLAEQGPFDVILHKITSKEWQQVLED YHEEHPEVTVLDPPNAINHLNNRQSMLAEVSDLNLSSFYGEVCTPRQLVIMRDPSSIP TAVAMAGLTLPLVAKPLVVDGTSKSHELSLAYDEASLSMLDPPLVLQEFVNHGGILFK VYIIGETIQVVRRFSLPDVNTYDLLNNVGVYRFPRVSCAAASADHADLDPHISELPPR PLLEKLGKELRGRLGLRLFNIDMIRELGTKDRYYIIDINYFPGFGKMPGYEHIFTDFL LNLAQSKYKKCLSGG</aaseq> |
| Gene Sequence |
<dnaseqindica>243..430#1845..1926#2113..2145#2445..2565#3336..3428#3715..3834#3934..4110#4395..4450#4523..4607#4904..4998#ttccactcgtctcgccattaacaagaattctttttttccttcccccaaatccttctcgagcaacagctaggggggagtggttcgcttcgtaagaagccggctacctctgctcgatcccgcctttgcctgcggattcccaccctgattttcgccgtttgtttgtttagtttcttggggcaaattcgagacgaggggaggagagtttttttgggggggagttgttgcggccttgcgggcggggtatgcggctgcacggggaggtttccttcgatgaggacgaggaggaagtggtgatggttccggcggcggcgctgtcgtcgtcgccgctcaacggcggggccgttccggtgacgaggctcgtggtggggtacgccctcaccaagaagaaggtgaagagcttccttcagcccaatctgctgttgctggcgaggtaaaaacctggatcaacgctcaaagagatgaacctgctgattcgtggccgtggtggaggattttgggagacaatagagtatttttgttggatttcttgttgatcgagtggcaacgaaatttttttcactaaaaccagaattttttgctgttggagggaaatggtaaatgatttgaggggggaatgcatggtttatgagatacagttttgcattttgagtctttgattaaatgctgtaattggttctatttctgaagcccagaggatttttgcgagaacaggtaaaaaacttggtagttccataacaaaatggggggtgggaatggaatgcaattctattccttgtagcactaatcaagtagtgatcgaatcgaacctctactggagtataaagttagcaggggcggctagtgatagtggattctgggcgaagcttgctgcatttgcgatatattgtaaactgaagaatatggctagtgtttattaggaattctggtagcatttgtagatgcccccttacgtgcgtctcatgttgacagttccactgcatgcgggggtctctatagtctataattgtatattatcatgccaacttgtcaaaaatgggtcttagcatgcaaatctttgctacttggtgtctgggattctagtttggaatattcccataatcccatggataagcacatcacatggattacgacagctttagtgatggcaatgccttttctgggatcgtttacaatttagtgactgacttgctaggtttgatggcaacatggaccgctgacaaacgatctttgtgattgtcttgttagctccctgaatcaagatccaagctgactagctgagtcaatcatttgctagttgctcggtaggctgcagacataagcaacacaagctgaaatatagaagggaaatgaaattaatatatgaaatacttatgcaaaatcaaaatgtttttgcttaaaactatttgtgtattgcatgcctagtttgtgcttttgtttcgatagatttattgtttagttattccaagaaaataaaggaaacaatatatatttatgcaaaatccaactatattcattcaaacaacctgcatattctctaaacgcttttttatatccctgtagtagtcttattttcaaagtataattgagaaatactctgttgcaaggcagtttaacttttttttatcattcttatttatactatcgatagagtataaatgtagcccaaacgacatgttatacatggagagatgatgatgtactaaaaaaattcttgattctgtgagggcttggacttggtcaagatggctattgtacttccatatggatgttaggatccatttctgtttcttgccattcttcttgtataacactatttcatgatgtttcattatgtactgtgtcaggaagaagggaattaatcttgtagcaattgatgacactcgcccgcttgcagaacaaggcccatttgatgttatcttgcacaaggttagtcgtgacctttttaaagctggactaaaatagtgctgcccattggccctgtatgatgcattttgttaaagaataacatcatctatttatgagctttagttataattaatgttttctgataattatgtcattgtaatttggtaaagtatatgcatattttattgaagaaatttctttgtgcagattactagcaaggaatggcaacaggttctggaggtaagatctgagtacgtatgcagaaatctgatgatgcacagtaaatgaccataccatcgttgccttttttctacttgttcactgcctcatcatggggaaatatgttatcaactgaggaaatgagcagtagatgtttgtcactgtatgtaatgattatatatccaaactatttgattgcagtgccctattgtaaagctcaacataaatccataattcttggaatttgctgttggcttctaatctgtgttgcccatttattgaaactctctgttaacctaactcctttcactttcctgaaggattatcatgaggaacatccagaggttactgtccttgacccaccaaatgccatcaatcatctgaataatcggcaatctatgcttgcagaagtatctgatttgaacttatccagtttctatggtaatcatgaagcttaacatgtatatttatatgctataaaatggattcgttattttactttccatttctaatgtttataatgttagttagtgtctgtttaattatcgaatatattcacatgtcagggaagcactggtcatgtggtgtagtcgtaggtactccacctgccagggttcaaatcctgatgctcacgaatattatgcacacccaagtgggctttcactagaaacttagtgagatcagcgattgtcgttggtttctgtctcttagagtgtgtgttaggggcgcatcagttgagttagtgtggtgcgcgcgcgtgtgcgtctgccgtgtagtcgcaaaaaagaaaaatatattcacatttaaacttctgtcctagaatggaagaaacttttgctttcaaaagcgatgtgcagaatatttatccttctaagataaacaatatatattttctattccttgtgaacctcaggattcaggaatttccaattaatgctgctatgtaattctttcttcaaacaaaagaaatttgtgttaactcttagaaatgctgaaattcccaatagaaacccatagatttttttgctttgcagagatttcaaggacaaatgttgcttgattgggaaacattatttctttgcagctggttgtgaatacaatgtaccgagatagtaaaaggcagaattccaaaagtgtatcatcaactaaatctattgagggcatatctattcataccaatatatgattgtttatgcattgaaccattgcaggagaagtttgcactccacgccaactggtcattatgagagatccatcctctataccaaccgcagttgccatggctggactaaccttgcccttgggtaattggtcataatctacatcttaaaactatccatttcctctagataagaaaacattgccaaatcatgcgaatgtttaaatacagcggaaaaatattcaacatttttattccggcaatattttttcacctggccattctcagttcgtttttgaacatgcggtctgtttttcagataatatttaagcatggttcgttgatagttaattttgggagatgttctcttttcattcttggaactattgagcttagaaattgtttgatgcgaagtgcttctgtctatacagttgcgaagccattggttgttgatggaacatctaaatcccacgaactatctcttgcatatgatgaggcatccttatcaatgcttgatcctcctctggtcctccaggaatttgtgaaccatggtaagttctgttccccccctgtaactgtagtacttttgcatatcctgagactttttcctccaacggtagaagttcggacctaaactattccctctacaggtgggatcctctttaaggtgtacatcattggtgaaactatacaagttgtacggaggttttctcttcctgatgttaacacttatgacttattaaacaatgttggcgtctatcgatttccaagagtttcatgtgctgcagctagtgcagaccatgcagacctcgatcctcatatctcaggtgagacatctgcagtattgtagttaatcttcattttggttattattgatccatgcttcattaatgtaagagggggcactaaattcaaacacaaactttctttagaaccttaacgcagtgtacatggtaaagagtttttatgtttccttaccatatatcaagtcgaggacagcgtttgttctatgacatcgatacttcttgatttattttgcttttgatgagtagtcgacggcaccagataaacttttgctatatattgatcagaactttctgttacatgatagaacttcctccaagaccactcctagagaaactgggaaaagagcttcgtggaagactggtatggccatgtatttatattaaccaagctctgtttcatatggtcgtcatctgacaggcttaaaaatttcagggtttaagattgttcaacatagatatgatcagagaacttggaaccaaagatcggtactatataattgacatcaactacttcccaggtgtgcagttattttcccactgccttcataaatggcatactatcatttgcgtttcttttcttctcttcatatgttcgtgcagaatttgttaggcctgattttattccagcctaaggggaaaggaggatacatcgctacacatacctggacttctaggccaaattaatactgatccagtttatccagtagtactttgtcgtctcgaatttctgtgttcttgttatttaacacagcatgatcattgttctgcagcagcattttcttgagtgtacgctcactccctcttgcttgcgcagggttcgggaaaatgccaggttatgagcacatattcaccgatttcttgctgaatcttgcgcaaagcaagtacaagaagtgcttaagcggcggctgaagtgcaaagagtcctgctgaacattaacaaaatggaatgtaacggtctagcagtcagtgtacatatctcggagaaataagtttaatcccaaggctttgaggaagagagatttagggtgtcttcccagaaaatggtggcacttaccggattagagaagagatgaaaaaatggatcgattttctaatgccgagtgcttgtataaatcactaacactgaatgttcccagcgtgccttcgtcaaggcttctgtctcaataatttcattatctttttcatgagcaatccttttagaatg</dnaseqindica> 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