Differential expression and subcellular localization of BmHMGA in silkworm (Bombyx mori)
The high mobility group A (HMGA) proteins have a close relation with chromatin remodeling and cancer transformation. They are widely expressed during embryogenesis, whereas their expression is absent or very low in adult tissues. The full open reading frame (ORF) of Bombyx mori high mobility group A...
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Zhejiang University Press
2014-05-01
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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.1008-9209.2013.12.011 |
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| author | Zhang Haihua Zhao Yan Zhang Hanming Li Si Quan Yanping Yu Wei Zhang Yaozhou Tong Fudan Zhang Xiaojuan |
| author_facet | Zhang Haihua Zhao Yan Zhang Hanming Li Si Quan Yanping Yu Wei Zhang Yaozhou Tong Fudan Zhang Xiaojuan |
| author_sort | Zhang Haihua |
| collection | DOAJ |
| description | The high mobility group A (HMGA) proteins have a close relation with chromatin remodeling and cancer transformation. They are widely expressed during embryogenesis, whereas their expression is absent or very low in adult tissues. The full open reading frame (ORF) of Bombyx mori high mobility group A (BmHMGA) was cloned with the cDNA of B. mori pupae, and the recombinant plasmid, pET-28a (+)-BmHMGA was constructed. The fusion protein His-BmHMGA was correctly expressed in BL21Star (DE3). New Zealand rabbits were immunized with fusion protein purified by N-i NTA affinity chromatography and the polyclonal antibody was obtained. The antibody had a high specificity and a titer of over 1∶12 800. The results of rea-l time PCR (RT-PCR) showed that the transcription levels of the BmHMGA gene declined with developmental stage in the order egg, fifth instar larva, pupa and moth. In the fifth instar larva stage, the transcription levels of the BmHMGA gene varied between tissues, declining in the order head, epidermis, ovaries, testis, fat body, stigma, Malpighian tubules, silk glands and midgut. The results of Western blotting and sem-i quantitative analysis by enzyme-linked immuno sorbent assay (ELISA) showed that the BmHMGA content was the highest in the egg and the lowest in the moth, and that in the fifth instar larva, the protein levels were obviously higher in the head than that in other tissues. No signals were detected in the midgut or Malpighian tubules. These results were identical to those of RT-PCR. The result of immunocytochemistry in BmN cells showed that the proteins were evenly distributed in the cytoplasm. |
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| institution | Kabale University |
| issn | 1008-9209 2097-5155 |
| language | English |
| publishDate | 2014-05-01 |
| 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-5ad7d92a466e4b4187d43ca565f417b02025-08-20T03:34:05ZengZhejiang University Press浙江大学学报. 农业与生命科学版1008-92092097-51552014-05-014023724810.3785/j.issn.1008-9209.2013.12.01110089209Differential expression and subcellular localization of BmHMGA in silkworm (Bombyx mori)Zhang HaihuaZhao YanZhang HanmingLi SiQuan YanpingYu WeiZhang YaozhouTong FudanZhang XiaojuanThe high mobility group A (HMGA) proteins have a close relation with chromatin remodeling and cancer transformation. They are widely expressed during embryogenesis, whereas their expression is absent or very low in adult tissues. The full open reading frame (ORF) of Bombyx mori high mobility group A (BmHMGA) was cloned with the cDNA of B. mori pupae, and the recombinant plasmid, pET-28a (+)-BmHMGA was constructed. The fusion protein His-BmHMGA was correctly expressed in BL21Star (DE3). New Zealand rabbits were immunized with fusion protein purified by N-i NTA affinity chromatography and the polyclonal antibody was obtained. The antibody had a high specificity and a titer of over 1∶12 800. The results of rea-l time PCR (RT-PCR) showed that the transcription levels of the BmHMGA gene declined with developmental stage in the order egg, fifth instar larva, pupa and moth. In the fifth instar larva stage, the transcription levels of the BmHMGA gene varied between tissues, declining in the order head, epidermis, ovaries, testis, fat body, stigma, Malpighian tubules, silk glands and midgut. The results of Western blotting and sem-i quantitative analysis by enzyme-linked immuno sorbent assay (ELISA) showed that the BmHMGA content was the highest in the egg and the lowest in the moth, and that in the fifth instar larva, the protein levels were obviously higher in the head than that in other tissues. No signals were detected in the midgut or Malpighian tubules. These results were identical to those of RT-PCR. The result of immunocytochemistry in BmN cells showed that the proteins were evenly distributed in the cytoplasm.https://www.academax.com/doi/10.3785/j.issn.1008-9209.2013.12.011<italic>Bombyx mori</italic>high mobility group Adifferential expressionsubcellular localizationimmunocytochemistry |
| spellingShingle | Zhang Haihua Zhao Yan Zhang Hanming Li Si Quan Yanping Yu Wei Zhang Yaozhou Tong Fudan Zhang Xiaojuan Differential expression and subcellular localization of BmHMGA in silkworm (Bombyx mori) 浙江大学学报. 农业与生命科学版 <italic>Bombyx mori</italic> high mobility group A differential expression subcellular localization immunocytochemistry |
| title | Differential expression and subcellular localization of BmHMGA in silkworm (Bombyx mori) |
| title_full | Differential expression and subcellular localization of BmHMGA in silkworm (Bombyx mori) |
| title_fullStr | Differential expression and subcellular localization of BmHMGA in silkworm (Bombyx mori) |
| title_full_unstemmed | Differential expression and subcellular localization of BmHMGA in silkworm (Bombyx mori) |
| title_short | Differential expression and subcellular localization of BmHMGA in silkworm (Bombyx mori) |
| title_sort | differential expression and subcellular localization of bmhmga in silkworm bombyx mori |
| topic | <italic>Bombyx mori</italic> high mobility group A differential expression subcellular localization immunocytochemistry |
| url | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2013.12.011 |
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