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《China Biotechnology》 2015-01
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Two Oleosins Flanking the KGF- 2 Improve the Expression Level of KGF-2 in Arabidopsis thaliana and Its Activity Analysis

AI Jun;JIANG Chao;LIU Min;WANG Xiao-yan;TIAN Hai-shan;LI Xiao-kun;Key Laboratory of Biotechnology Pharmaceutical Engineering of Zhejiang Province,School of Pharmaceutical Science,Wenzhou Medical University;Engineering Research Center of Bioreactor and Pharmaceutical Development,Ministry of Education;  
Objective: Construct the expression vector p1301-YO-K2-YO based on the oil body expression technology which has two oleosins flanking the KGF-2,observe whether it can improve the expression level of KGF-2 and identify its biological activity. Methods: KGF-2 core gene and Arabidopsis oleosin gene were amplified by PCR,KGF-2-oleosin fused sequence was obtained by the fusion PCR,ligated with the blank vector also has a oleosin gene( p1301-oleosin),then named it p1301-YO-K2-YO,which can be tanslated into the oleosin-KGF-2-oleosin fusion protein. Agrobacterium-mediated transformation of Arabidopsis thaliana was conducted by floral dip. The total protein of the T2 seeds were extracted for SDS-PAGE,Western blot analysis,the proliferation activity of hair follicles was tested on C57 BL /6 mice. Results: The plasmid which has two oleosins flanking the KGF-2 was successfully expressed in Arabidopsis thaliana. Western blot result shows this fusion expression strategy could be twice more higher than the single-oleosin configuration production in KGF-2 protein expression,and has a certain biological activity.
【Fund】: 浙江省自然科学基金(LY13H300006);; 2012年省级重点科技创新团队项目(2012R1004210);; 吉林省科技型中小企业创新基金(13C26212201223)资助项目
【CateGory Index】: Q946
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