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

Abstract

Adiponectin is a polypeptide specifi cally secreted from human adipocytes, and its defi ciency 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 modifi ed 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 amplifi cation promoting sequence acting as an enhancer of transcription was also introduced to enhance gene expression. The presence of the transgene and mRNA transcripts was confi rmed 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 modifi ed 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.

A novel synthetic Cry1Ab gene resists rice insect pests

“…Transgenic technology can break reproductive isolation to realize gene transfer among different species, and rice is a potential plant that can be used as a recombinant protein expres- sion system (Liu et al, 2011;Kim et al, 2012;An et al, 2013;Kuo et al, 2013). However, after a high-expression gene in one species it is transferred to another species, the expression is often reduced greatly, particularly for species with a large expression difference, leading to the loss of the gene's function (Gustafsson et al, 2004).…”