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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Autores principales:	Liu,Feng, Zhao,Yi-Ying, Zheng,Jin-Gui
Lenguaje:	English
Publicado:	Sociedad de Biología de Chile 2011
Materias:	human adiponectin transgenic rice modified gene anti-diabetic
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602011000400009
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spelling	oai:scielo:S0716-976020110004000092012-02-10High-level expression of modified gene encoding human adiponectin in transgenic riceLiu,FengZhao,Yi-YingZheng,Jin-Gui human adiponectin transgenic rice modified gene anti-diabetic 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.info:eu-repo/semantics/openAccessSociedad de Biología de ChileBiological Research v.44 n.4 20112011-01-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602011000400009en10.4067/S0716-97602011000400009
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	human adiponectin transgenic rice modified gene anti-diabetic
spellingShingle	human adiponectin transgenic rice modified gene anti-diabetic Liu,Feng Zhao,Yi-Ying Zheng,Jin-Gui High-level expression of modified gene encoding human adiponectin in transgenic rice
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.
author	Liu,Feng Zhao,Yi-Ying Zheng,Jin-Gui
author_facet	Liu,Feng Zhao,Yi-Ying Zheng,Jin-Gui
author_sort	Liu,Feng
title	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_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_sort	high-level expression of modified gene encoding human adiponectin in transgenic rice
publisher	Sociedad de Biología de Chile
publishDate	2011
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602011000400009
work_keys_str_mv	AT liufeng highlevelexpressionofmodifiedgeneencodinghumanadiponectinintransgenicrice AT zhaoyiying highlevelexpressionofmodifiedgeneencodinghumanadiponectinintransgenicrice AT zhengjingui highlevelexpressionofmodifiedgeneencodinghumanadiponectinintransgenicrice
_version_	1718441481496166400

High-level expression of modified gene encoding human adiponectin in transgenic rice

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