Prokaryotic and tissue-specific expression of RglKAT gene from Rehmannia glutinosa
A 1640-bp full-length cDNA of the 3-ketoacyl-CoA thiolase gene was cloned from Rehmannia glutinosa L. leaf using RT-PCR and RACE. Bioinformatics analyses indicated that it contained one 1395-bp open reading frameencoding a thiolase of 464 amino acids with the predicted molecular weight of 49 kDa, an...
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Indian Council of Agricultural Research
2015-11-01
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| Series: | The Indian Journal of Agricultural Sciences |
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| Online Access: | https://epubs.icar.org.in/index.php/IJAgS/article/view/53762 |
| _version_ | 1797771264290455552 |
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| author | Y Q ZHOU Y H ZHANG W S WANG X N WANG H Y DUAN |
| author_facet | Y Q ZHOU Y H ZHANG W S WANG X N WANG H Y DUAN |
| author_sort | Y Q ZHOU |
| collection | DOAJ |
| description | A 1640-bp full-length cDNA of the 3-ketoacyl-CoA thiolase gene was cloned from Rehmannia glutinosa L. leaf using RT-PCR and RACE. Bioinformatics analyses indicated that it contained one 1395-bp open reading frameencoding a thiolase of 464 amino acids with the predicted molecular weight of 49 kDa, and had a similarity of 73-82% to other species at the nucleotide level and 63-88% to other species at the amino acid level, respectively. The recombinant RglKAT was expressed in a bacterial system in an active form, analyzed by PAGE-CBB staining and purified using Ni-agarose chromatography. Real Time quantitative PCR analysis showed that relative expression levels of the RglKAT
gene differed between R.glutinosa tissues, with the highest levels in the petals and the lowest in the early senescing stems. These results revealed that RglKAT gene could be cloned and heterogeneously expressed, and related to R. glutinosa growth and development. |
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| issn | 0019-5022 2394-3319 |
| language | English |
| last_indexed | 2024-03-12T21:35:01Z |
| publishDate | 2015-11-01 |
| publisher | Indian Council of Agricultural Research |
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| series | The Indian Journal of Agricultural Sciences |
| spelling | doaj.art-ef21f4b918f346ebb1b0478ea6c0c4d42023-07-27T11:41:09ZengIndian Council of Agricultural ResearchThe Indian Journal of Agricultural Sciences0019-50222394-33192015-11-01851110.56093/ijas.v85i11.53762Prokaryotic and tissue-specific expression of RglKAT gene from Rehmannia glutinosaY Q ZHOU0Y H ZHANG1W S WANG2X N WANGH Y DUAN3Professor, College of Life Sciences, Henan Normal University, Xinxiang Henan, 453 007, China; and Key Laboratory for Microorganisms and Functional Molecules, University of Henan Province, No. 46 Jianshe Road, Xinxiang Henan, 453 007, P R ChinaGraduates, Plant Genetics, College of Life Science, College of Life Sciences, Henan Normal University, Xinxiang Henan, 453 007, China; and Key Laboratory for Microorganisms and Functional Molecules, University of Henan Province, No. 46 Jianshe Road, Xinxiang Henan, 453 007, P R ChinaCollege of Life Sciences, Henan Normal University, Xinxiang Henan, 453 007, China; and Key Laboratory for Microorganisms and Functional Molecules, University of Henan Province, No. 46 Jianshe Road, Xinxiang Henan, 453 007, P R ChinaCollege of Life Sciences, Henan Normal University, Xinxiang Henan, 453 007, China; and Key Laboratory for Microorganisms and Functional Molecules, University of Henan Province, No. 46 Jianshe Road, Xinxiang Henan, 453 007, P R ChinaA 1640-bp full-length cDNA of the 3-ketoacyl-CoA thiolase gene was cloned from Rehmannia glutinosa L. leaf using RT-PCR and RACE. Bioinformatics analyses indicated that it contained one 1395-bp open reading frameencoding a thiolase of 464 amino acids with the predicted molecular weight of 49 kDa, and had a similarity of 73-82% to other species at the nucleotide level and 63-88% to other species at the amino acid level, respectively. The recombinant RglKAT was expressed in a bacterial system in an active form, analyzed by PAGE-CBB staining and purified using Ni-agarose chromatography. Real Time quantitative PCR analysis showed that relative expression levels of the RglKAT gene differed between R.glutinosa tissues, with the highest levels in the petals and the lowest in the early senescing stems. These results revealed that RglKAT gene could be cloned and heterogeneously expressed, and related to R. glutinosa growth and development.https://epubs.icar.org.in/index.php/IJAgS/article/view/53762CloningExpression3-Ketoacyl-CoA thiolase (KAT)Rehmannia glutinosaReal Time quantitative PCR |
| spellingShingle | Y Q ZHOU Y H ZHANG W S WANG X N WANG H Y DUAN Prokaryotic and tissue-specific expression of RglKAT gene from Rehmannia glutinosa The Indian Journal of Agricultural Sciences Cloning Expression 3-Ketoacyl-CoA thiolase (KAT) Rehmannia glutinosa Real Time quantitative PCR |
| title | Prokaryotic and tissue-specific expression of RglKAT gene from Rehmannia glutinosa |
| title_full | Prokaryotic and tissue-specific expression of RglKAT gene from Rehmannia glutinosa |
| title_fullStr | Prokaryotic and tissue-specific expression of RglKAT gene from Rehmannia glutinosa |
| title_full_unstemmed | Prokaryotic and tissue-specific expression of RglKAT gene from Rehmannia glutinosa |
| title_short | Prokaryotic and tissue-specific expression of RglKAT gene from Rehmannia glutinosa |
| title_sort | prokaryotic and tissue specific expression of rglkat gene from rehmannia glutinosa |
| topic | Cloning Expression 3-Ketoacyl-CoA thiolase (KAT) Rehmannia glutinosa Real Time quantitative PCR |
| url | https://epubs.icar.org.in/index.php/IJAgS/article/view/53762 |
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