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Acta Agron Sin ›› 2015, Vol. 41 ›› Issue (04): 524-530.doi: 10.3724/SP.J.1006.2015.00524

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Enhancing Expression and Accumulation of Foreign Proteins by Using the Signal Peptide of Glutelin GluA-2 in Endosperm of Transgenic Rice

WANG Hong-Mei1,ZHANG Chang-Quan1,LI Qian-Feng1, SUN Samuel Sing-Min2,LIU Qiao-Quan1,*,XU Ming-Liang1,3   

  1. 1 Jiangsu Key Laboratory for Crop Genetics and Physiology / Key Laboratory of Plant Functional Genomics of Ministry of Education /Co-Innovation Center for Modern Production Technology of Grain Crops, Yangzhou University, Yangzhou 225009, China; 2 Department of Biology,The Chinese University of Hong Kong, Hong Kong, China; 3 College of Agriculture and Biotechnology, Chinese Agricultural University, Beijing 100193, China
  • Received:2014-11-01 Revised:2015-02-06 Online:2015-04-12 Published:2015-03-03

Abstract:

It is one of the key important techniques to enhance the expression of foreign proteins in target tissue/organ of transgenic plants. Glutelin is the major component of storage proteins in rice seeds, and its expression was tightly temporal and tissuespecific, which is controlled by several mechanisms. To further reveal the function of the Glutelin signal peptide on expression of target gene, in present study, we isolated the promoter and signal peptide-coding sequences of the glutelin GluA-2 gene, and fused them transcriptionally to the GUS coding sequences. Beside, the construct without the GluA-2 signal peptide-coding sequences was also generated as a control. Both constructs with the GUS chimeric genes, named as p13GSG and p13GG, were introduced into the same rice variety by Agrobacterium-mediated transformation. More than twenty independent transgenic lines were generated for each construct, and the integration of the GUS chimeric gene was confirmed by PCR technique. The results from Northern blot analysis showed that, after fusing the GluA-2 signal peptide coding sequences between the GluA-2 promoter and the GUS coding sequence, the transcription of GUS chimeric gene could be dramatically increased. Then, Western blot was carried out by using the GUS-specific antibody, and the results obviously revealed that the accumulation of foreign proteins was significantly
enhanced in the endosperm of transgenic rice with the signal peptide. However, there was no or very low GUS activity in the endosperm of transgenic rice plants with the signal peptide. These results were very useful to improve the grain quality of rice via genetic engineering, especially produce foreign proteins in the seeds of rice as bioreactor.

Key words: Transgenic rice, Signal peptide, Glutelin, Endosperm, Gene expression

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