Chinese Journal OF Rice Science >
Construction of Seed Protein Body Targeted Expression Vector and Transformation Rice for Oral Recombinant Proinsulin
*Corresponding author：E-mailyanzhao9918@163.com
Received date: 2014-12-28
Revised date: 2015-03-25
Online published: 2015-09-10
Developent of oral recombinant proinsulin using rice seeds as bioreactor shows good application future. A fusion gene of the cholera toxin B subunit and human proinsulin (CTBIN) with an endoplasmic reticulum retention signal (KDEL) at C-terminus was designed for its self-maturation process in human gut and synthesized according to the optimized code usage bias of rice. The synthetic CTBIN was introduced to the vector pCAMBIA1302 and expressed under the 2.4 kb promoter sequence of rice glutelin GluB1 with its signal peptide (pGluB1sig), which was cloned by PCR from the japonica rice Nipponbare genome. Thus the rice seed protein body targeted expression vector pCAMBIA1302-pGluB1sig-CTBIN-Nos for oral delivery of recombinant proinsulin was constructed and then transformed into Nipponbare via Agrobacterium-mediated method. Totally 46 transgenic rice plants were obtained and the expression of the fusion protein CTB::Human proinsulin in rice seeds was detected by Western-blot.
Yong-zhou TANG, Yue YU, Yan ZHAO . Construction of Seed Protein Body Targeted Expression Vector and Transformation Rice for Oral Recombinant Proinsulin[J]. Chinese Journal OF Rice Science, 2015 , 29(5) : 481 -489 . DOI: 10.3969/j.issn.1001G7216.2015.05.005
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