作物学报Issue(4)：524-530,7.DOI:10.3724/SP.J.1006.2015.00524
以谷蛋白GluA-2信号肽增强外源蛋白在转基因水稻胚乳中的表达与积累
Enhancing Expression and Accumulation of Foreign Proteins by Using the Signal Peptide of Glutelin GluA-2 in Endosperm of Transgenic Rice
摘要
Abstract
It is one of the key important techniques to enhance the expression of foreign proteins in target tissue/organ of trans-genic plants. Glutelin is the major component of storage proteins in rice seeds, and its expression was tightly temporal and tissue- specific, 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 glutelinGluA-2 gene, and fused them transcriptionally to theGUS coding sequences. Beside, the construct without theGluA-2 signal peptide-coding sequences was also generated as a control. Both constructs with theGUS chimeric genes, named as p13GSG and p13GG, were introduced into the same rice variety byAgrobacterium-mediated transformation. More than twenty independent transgenic lines were gener-ated for each construct, and the integration of theGUS chimeric gene was confirmed by PCR technique. The results from North-ern blot analysis showed that, after fusing theGluA-2 signal peptide coding sequences between theGluA-2 promoter and theGUS coding sequence, the transcription ofGUS 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.关键词
转基因水稻/信号肽/谷蛋白/胚乳/基因表达Key words
Transgenic rice/Signal peptide/Glutelin/Endosperm/Gene expression引用本文复制引用
王红梅,张昌泉,李钱峰,辛世文,刘巧泉,徐明良..以谷蛋白GluA-2信号肽增强外源蛋白在转基因水稻胚乳中的表达与积累[J].作物学报,2015,(4):524-530,7.基金项目
本研究由国家转基因生物新品种培育重大专项(2014ZX08001006-005,2014ZX08009-024B),江苏省杰出青年基金项目(BK2012010),江苏省高校优势学科建设工程和“青蓝工程”等项目资助。 （2014ZX08001006-005,2014ZX08009-024B）
