High-level expression of modified gene encoding human adiponectin in transgenic rice
Adiponectin is a polypeptide specifically secreted from human adipocytes, and its deficiency is closely linked to increased obesity and type II diabetes. There is an urgent demand for large-scale production of human adiponectin for pharmaceutical applications. Here, we report that we have successful...
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| Format: | Article |
| Language: | English |
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BMC
2011-01-01
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| Series: | Biological Research |
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| Online Access: | http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602011000400009 |
| _version_ | 1818232030679269376 |
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| author | Feng Liu Yi-Ying Zhao Jin-Gui Zheng |
| author_facet | Feng Liu Yi-Ying Zhao Jin-Gui Zheng |
| author_sort | Feng Liu |
| collection | DOAJ |
| description | Adiponectin is a polypeptide specifically secreted from human adipocytes, and its deficiency is closely linked to increased obesity and type II diabetes. There is an urgent demand for large-scale production of human adiponectin for pharmaceutical applications. Here, we report that we have successfully obtained a high-level of expression of modified genes encoding human adiponectin in transgenic rice. The 735 bp cDNA of the native human sequence was adopted to rice codon usage, fused to the translation initiation sequence in the N terminus and to the KDEL signal sequence in the C terminus. An amplification promoting sequence acting as an enhancer of transcription was also introduced to enhance gene expression. The presence of the transgene and mRNA transcripts was confirmed by PCR, Southern blot and RT-PCR. Western blot analysis revealed that a protein of approximately 30 kDa was produced in rice leaves. ELISA analysis was used to determine the amount of recombinant adiponectin in transformants with the modified gene in up to 0.32% of total soluble leaf protein. Our results establish the feasibility of high-level expression of recombinant human adiponectin in transgenic rice. |
| first_indexed | 2024-12-12T10:59:48Z |
| format | Article |
| id | doaj.art-9e7a65508ac3462381c4a73188f43a69 |
| institution | Directory Open Access Journal |
| issn | 0716-9760 0717-6287 |
| language | English |
| last_indexed | 2024-12-12T10:59:48Z |
| publishDate | 2011-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Biological Research |
| spelling | doaj.art-9e7a65508ac3462381c4a73188f43a692022-12-22T00:26:35ZengBMCBiological Research0716-97600717-62872011-01-01444369375High-level expression of modified gene encoding human adiponectin in transgenic riceFeng LiuYi-Ying ZhaoJin-Gui ZhengAdiponectin is a polypeptide specifically secreted from human adipocytes, and its deficiency is closely linked to increased obesity and type II diabetes. There is an urgent demand for large-scale production of human adiponectin for pharmaceutical applications. Here, we report that we have successfully obtained a high-level of expression of modified genes encoding human adiponectin in transgenic rice. The 735 bp cDNA of the native human sequence was adopted to rice codon usage, fused to the translation initiation sequence in the N terminus and to the KDEL signal sequence in the C terminus. An amplification promoting sequence acting as an enhancer of transcription was also introduced to enhance gene expression. The presence of the transgene and mRNA transcripts was confirmed by PCR, Southern blot and RT-PCR. Western blot analysis revealed that a protein of approximately 30 kDa was produced in rice leaves. ELISA analysis was used to determine the amount of recombinant adiponectin in transformants with the modified gene in up to 0.32% of total soluble leaf protein. Our results establish the feasibility of high-level expression of recombinant human adiponectin in transgenic rice.http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602011000400009human adiponectintransgenic ricemodified geneanti-diabetic |
| spellingShingle | Feng Liu Yi-Ying Zhao Jin-Gui Zheng High-level expression of modified gene encoding human adiponectin in transgenic rice Biological Research human adiponectin transgenic rice modified gene anti-diabetic |
| title | High-level expression of modified gene encoding human adiponectin in transgenic rice |
| title_full | High-level expression of modified gene encoding human adiponectin in transgenic rice |
| title_fullStr | High-level expression of modified gene encoding human adiponectin in transgenic rice |
| title_full_unstemmed | High-level expression of modified gene encoding human adiponectin in transgenic rice |
| title_short | High-level expression of modified gene encoding human adiponectin in transgenic rice |
| title_sort | high level expression of modified gene encoding human adiponectin in transgenic rice |
| topic | human adiponectin transgenic rice modified gene anti-diabetic |
| url | http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602011000400009 |
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