Cloning and expression analysis of pathogenesis-related protein 1 gene in maize
By reverse transcription-polymerase chain reaction (RT-PCR), an open reading frame of pathogenesis-related protein 1 (PR1) was isolated from inbred line R15 which was resistant to kernel rot, named ZmPR1. Molecular and bioinformatic analyses of ZmPR1 revealed that an open reading frame of 528 bp was...
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Zhejiang University Press
2012-01-01
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| Series: | 浙江大学学报. 农业与生命科学版 |
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| Online Access: | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2012.01.005 |
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| author | WANG Jing LIU Li ZHANG Zhi-ming ZHAO Mao-jun PAN Guang-tang |
| author_facet | WANG Jing LIU Li ZHANG Zhi-ming ZHAO Mao-jun PAN Guang-tang |
| author_sort | WANG Jing |
| collection | DOAJ |
| description | By reverse transcription-polymerase chain reaction (RT-PCR), an open reading frame of pathogenesis-related protein 1 (PR1) was isolated from inbred line R15 which was resistant to kernel rot, named ZmPR1. Molecular and bioinformatic analyses of ZmPR1 revealed that an open reading frame of 528 bp was predicted to encode a 175-amino acid protein with a deduced molecular mass of 18.7 ku. Homology analysis showed that the deduced amino acid sequence of PR1 protein in maize had a high similarity with other higher plants, such as Oryza sativa, Triticum aestivum, and Arabidopsis. PR1 proteins from different plants had the same conservative structure domain of cysteine-rich secretory protein (CAP). The recombinant expressed plasmid pET32a(+)-ZmPR1 was expressed in Escherichia coli BL21. The expression conditions were optimized by induction at different time, different temperature, and different IPTG concentrations. The results indicated that the expression products migrated at the size of 39 ku by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis. The optimum expression condition was 0.6 mmol·L<sup>-1</sup> IPTG at 28 ℃, but the induction time has little effect on the expression. The successful expression of ZmPR1 provides some basis for protein purification and preparation of the monoclonal antibody. |
| format | Article |
| id | doaj-art-41c8e1d21ee04ff59c178ed48e98f6b6 |
| institution | Matheson Library |
| issn | 1008-9209 2097-5155 |
| language | English |
| publishDate | 2012-01-01 |
| publisher | Zhejiang University Press |
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| series | 浙江大学学报. 农业与生命科学版 |
| spelling | doaj-art-41c8e1d21ee04ff59c178ed48e98f6b62025-08-01T05:33:24ZengZhejiang University Press浙江大学学报. 农业与生命科学版1008-92092097-51552012-01-0138354210.3785/j.issn.1008-9209.2012.01.00510089209Cloning and expression analysis of pathogenesis-related protein 1 gene in maizeWANG JingLIU LiZHANG Zhi-mingZHAO Mao-junPAN Guang-tangBy reverse transcription-polymerase chain reaction (RT-PCR), an open reading frame of pathogenesis-related protein 1 (PR1) was isolated from inbred line R15 which was resistant to kernel rot, named ZmPR1. Molecular and bioinformatic analyses of ZmPR1 revealed that an open reading frame of 528 bp was predicted to encode a 175-amino acid protein with a deduced molecular mass of 18.7 ku. Homology analysis showed that the deduced amino acid sequence of PR1 protein in maize had a high similarity with other higher plants, such as Oryza sativa, Triticum aestivum, and Arabidopsis. PR1 proteins from different plants had the same conservative structure domain of cysteine-rich secretory protein (CAP). The recombinant expressed plasmid pET32a(+)-ZmPR1 was expressed in Escherichia coli BL21. The expression conditions were optimized by induction at different time, different temperature, and different IPTG concentrations. The results indicated that the expression products migrated at the size of 39 ku by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis. The optimum expression condition was 0.6 mmol·L<sup>-1</sup> IPTG at 28 ℃, but the induction time has little effect on the expression. The successful expression of ZmPR1 provides some basis for protein purification and preparation of the monoclonal antibody.https://www.academax.com/doi/10.3785/j.issn.1008-9209.2012.01.005maize kernel rotpathogenesis-related proteinprokaryotic expressionexpression analysis |
| spellingShingle | WANG Jing LIU Li ZHANG Zhi-ming ZHAO Mao-jun PAN Guang-tang Cloning and expression analysis of pathogenesis-related protein 1 gene in maize 浙江大学学报. 农业与生命科学版 maize kernel rot pathogenesis-related protein prokaryotic expression expression analysis |
| title | Cloning and expression analysis of pathogenesis-related protein 1 gene in maize |
| title_full | Cloning and expression analysis of pathogenesis-related protein 1 gene in maize |
| title_fullStr | Cloning and expression analysis of pathogenesis-related protein 1 gene in maize |
| title_full_unstemmed | Cloning and expression analysis of pathogenesis-related protein 1 gene in maize |
| title_short | Cloning and expression analysis of pathogenesis-related protein 1 gene in maize |
| title_sort | cloning and expression analysis of pathogenesis related protein 1 gene in maize |
| topic | maize kernel rot pathogenesis-related protein prokaryotic expression expression analysis |
| url | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2012.01.005 |
| work_keys_str_mv | AT wangjing cloningandexpressionanalysisofpathogenesisrelatedprotein1geneinmaize AT liuli cloningandexpressionanalysisofpathogenesisrelatedprotein1geneinmaize AT zhangzhiming cloningandexpressionanalysisofpathogenesisrelatedprotein1geneinmaize AT zhaomaojun cloningandexpressionanalysisofpathogenesisrelatedprotein1geneinmaize AT panguangtang cloningandexpressionanalysisofpathogenesisrelatedprotein1geneinmaize |