The isolation and expression analysis of a class I chitinase from developing winged bean seed (Psophocarpus tetragonolubus)
Chitinase catalyses the hydrolysis of β-1,4-N-acetyl-D-glucosamine linkages of the fungal cell wall polymer chitin and is involved in the inducible defenses of plants. The aim of this research was to isolate and clone chitinase cDNAs from the seed of winged bean. Chitinase gene fragments were isolat...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Malaysian Society for Molecular Biology and Biotechnology
2003
|
| Online Access: | http://psasir.upm.edu.my/id/eprint/58523/1/The%20isolation%20and%20expression%20analysis%20of%20a%20class%20I%20chitinase%20from%20developing%20winged%20bean%20seed%20%28Psophocarpus%20tetragonolubus%29.pdf |
| _version_ | 1796976917852717056 |
|---|---|
| author | Sekeli, Rogayah Tan, Yvonne Yik Wan Kulaveerasingam, Harikrishna Napis, Suhaimi Tan, Siang Hee |
| author_facet | Sekeli, Rogayah Tan, Yvonne Yik Wan Kulaveerasingam, Harikrishna Napis, Suhaimi Tan, Siang Hee |
| author_sort | Sekeli, Rogayah |
| collection | UPM |
| description | Chitinase catalyses the hydrolysis of β-1,4-N-acetyl-D-glucosamine linkages of the fungal cell wall polymer chitin and is involved in the inducible defenses of plants. The aim of this research was to isolate and clone chitinase cDNAs from the seed of winged bean. Chitinase gene fragments were isolated from a winged bean seed cDNA library using two sets of degenerate primers corresponding to the conserved regions of chitinase class I and IV. The poly A+ mRNA was reversed transcribed and further amplified using RT-PCR. A 1.1 Kb fragment was selected, cloned and sequenced. A nucleotide sequence comparison identified the fragment as a Class I basic chitinase cDNA; this fragment was subsequently used as a probe to screen for a full length transcript from the cDNA library. Library screening resulted in the isolation of a 1324 bp clone designated CHRZP; encoding a polypeptide of 289 amino acids containing the diagnostic N-terminal cysteine-rich domain of class 1 chitinases. CHRZP showed 47% similarity to a chitinase sequence from Rice and to another class I winged bean chitinase (ChitiWb1) at the amino acid level. RNA blot hybridization revealed that CHRZP mRNA accumulates to the highest level in leaves followed by tubers and pods. Southern hybridization analysis indicated that this gene is likely to be present as a single copy in the winged bean genome. |
| first_indexed | 2024-03-06T09:32:38Z |
| format | Article |
| id | upm.eprints-58523 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T09:32:38Z |
| publishDate | 2003 |
| publisher | Malaysian Society for Molecular Biology and Biotechnology |
| record_format | dspace |
| spelling | upm.eprints-585232018-01-22T02:24:57Z http://psasir.upm.edu.my/id/eprint/58523/ The isolation and expression analysis of a class I chitinase from developing winged bean seed (Psophocarpus tetragonolubus) Sekeli, Rogayah Tan, Yvonne Yik Wan Kulaveerasingam, Harikrishna Napis, Suhaimi Tan, Siang Hee Chitinase catalyses the hydrolysis of β-1,4-N-acetyl-D-glucosamine linkages of the fungal cell wall polymer chitin and is involved in the inducible defenses of plants. The aim of this research was to isolate and clone chitinase cDNAs from the seed of winged bean. Chitinase gene fragments were isolated from a winged bean seed cDNA library using two sets of degenerate primers corresponding to the conserved regions of chitinase class I and IV. The poly A+ mRNA was reversed transcribed and further amplified using RT-PCR. A 1.1 Kb fragment was selected, cloned and sequenced. A nucleotide sequence comparison identified the fragment as a Class I basic chitinase cDNA; this fragment was subsequently used as a probe to screen for a full length transcript from the cDNA library. Library screening resulted in the isolation of a 1324 bp clone designated CHRZP; encoding a polypeptide of 289 amino acids containing the diagnostic N-terminal cysteine-rich domain of class 1 chitinases. CHRZP showed 47% similarity to a chitinase sequence from Rice and to another class I winged bean chitinase (ChitiWb1) at the amino acid level. RNA blot hybridization revealed that CHRZP mRNA accumulates to the highest level in leaves followed by tubers and pods. Southern hybridization analysis indicated that this gene is likely to be present as a single copy in the winged bean genome. Malaysian Society for Molecular Biology and Biotechnology 2003 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/58523/1/The%20isolation%20and%20expression%20analysis%20of%20a%20class%20I%20chitinase%20from%20developing%20winged%20bean%20seed%20%28Psophocarpus%20tetragonolubus%29.pdf Sekeli, Rogayah and Tan, Yvonne Yik Wan and Kulaveerasingam, Harikrishna and Napis, Suhaimi and Tan, Siang Hee (2003) The isolation and expression analysis of a class I chitinase from developing winged bean seed (Psophocarpus tetragonolubus). Asia Pacific Journal of Molecular Biology and Biotechnology, 11 (2). pp. 63-69. ISSN 0128-7451 http://www.msmbb.org.my/apjmbb/html112/112.htm |
| spellingShingle | Sekeli, Rogayah Tan, Yvonne Yik Wan Kulaveerasingam, Harikrishna Napis, Suhaimi Tan, Siang Hee The isolation and expression analysis of a class I chitinase from developing winged bean seed (Psophocarpus tetragonolubus) |
| title | The isolation and expression analysis of a class I chitinase from developing winged bean seed (Psophocarpus tetragonolubus) |
| title_full | The isolation and expression analysis of a class I chitinase from developing winged bean seed (Psophocarpus tetragonolubus) |
| title_fullStr | The isolation and expression analysis of a class I chitinase from developing winged bean seed (Psophocarpus tetragonolubus) |
| title_full_unstemmed | The isolation and expression analysis of a class I chitinase from developing winged bean seed (Psophocarpus tetragonolubus) |
| title_short | The isolation and expression analysis of a class I chitinase from developing winged bean seed (Psophocarpus tetragonolubus) |
| title_sort | isolation and expression analysis of a class i chitinase from developing winged bean seed psophocarpus tetragonolubus |
| url | http://psasir.upm.edu.my/id/eprint/58523/1/The%20isolation%20and%20expression%20analysis%20of%20a%20class%20I%20chitinase%20from%20developing%20winged%20bean%20seed%20%28Psophocarpus%20tetragonolubus%29.pdf |
| work_keys_str_mv | AT sekelirogayah theisolationandexpressionanalysisofaclassichitinasefromdevelopingwingedbeanseedpsophocarpustetragonolubus AT tanyvonneyikwan theisolationandexpressionanalysisofaclassichitinasefromdevelopingwingedbeanseedpsophocarpustetragonolubus AT kulaveerasingamharikrishna theisolationandexpressionanalysisofaclassichitinasefromdevelopingwingedbeanseedpsophocarpustetragonolubus AT napissuhaimi theisolationandexpressionanalysisofaclassichitinasefromdevelopingwingedbeanseedpsophocarpustetragonolubus AT tansianghee theisolationandexpressionanalysisofaclassichitinasefromdevelopingwingedbeanseedpsophocarpustetragonolubus AT sekelirogayah isolationandexpressionanalysisofaclassichitinasefromdevelopingwingedbeanseedpsophocarpustetragonolubus AT tanyvonneyikwan isolationandexpressionanalysisofaclassichitinasefromdevelopingwingedbeanseedpsophocarpustetragonolubus AT kulaveerasingamharikrishna isolationandexpressionanalysisofaclassichitinasefromdevelopingwingedbeanseedpsophocarpustetragonolubus AT napissuhaimi isolationandexpressionanalysisofaclassichitinasefromdevelopingwingedbeanseedpsophocarpustetragonolubus AT tansianghee isolationandexpressionanalysisofaclassichitinasefromdevelopingwingedbeanseedpsophocarpustetragonolubus |