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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2015, Vol. 35 Issue (1): 21-26    DOI: 10.13523/j.cb.20150103
    
Two Oleosins Flanking the KGF-2 Improve the Expression Level of KGF-2 in Arabidopsis thaliana and Its Activity Analysis
AI Jun1, JIANG Chao1,2, LIU Min1,2, WANG Xiao-yan1, TIAN Hai-shan1, LI Xiao-kun1,2
1. Key Laboratory of Biotechnology Pharmaceutical Engineering of Zhejiang Province, School of Pharmaceutical Science, Wenzhou Medical University, Wenzhou 325035, China;
2. Engineering Research Center of Bioreactor and Pharmaceutical Development, Ministry of Education, Changchun 130118, China
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Abstract  

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 C57BL /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.

Key words： Arabidopsis thaliana      Keratinocyte growth factor-2      Oleosin-KGF-2-oleosin fusion protein     
Received: 19 October 2014      Published: 25 January 2015
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Cite this article:

AI Jun, JIANG Chao, LIU Min, WANG Xiao-yan, TIAN Hai-shan, LI Xiao-kun. Two Oleosins Flanking the KGF-2 Improve the Expression Level of KGF-2 in Arabidopsis thaliana and Its Activity Analysis. China Biotechnology, 2015, 35(1): 21-26.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20150103     OR     https://manu60.magtech.com.cn/biotech/Y2015/V35/I1/21


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