The codon-optimized capsid gene of duck circovirus can be highly expressed in yeast and self-assemble into virus-like particles

doi:10.1016/S2095-3119(16)61605-2

Journal of Integrative Agriculture ›› 2017, Vol. 16 ›› Issue (07): 1601-1608.DOI: 10.1016/S2095-3119(16)61605-2

收稿日期:2016-11-08 出版日期:2017-07-20 发布日期:2017-07-06

The codon-optimized capsid gene of duck circovirus can be highly expressed in yeast and self-assemble into virus-like particles

YANG Cui^{1, 2, 3*}, XU Yu^{1, 2, 3*}, JIA Ren-yong^{1, 2, 3*}, LIU Si-yang^{1, 2, 3*}, WANG Ming-shu^{1, 2, 3}, ZHU De-kang^{1, 3}, CHEN Shun^{1, 2, 3}, LIU Ma-feng^{1, 2, 3}, ZHAO Xin-xin^{1, 2, 3}, SUN Kun-feng^{1, 2, 3}, JING Bo³, YIN Zhong-qiong³, CHENG An-chun^{1, 2, 3}

¹ Avian Disease Research Centre, Sichuan Agricultural University, Chengdu 611130, P.R.China
² Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, P.R.China
³ Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu 611130, P.R.China

Received:2016-11-08 Online:2017-07-20 Published:2017-07-06
Contact: Correspondence JIA Ren-yong, Tel/Fax: +86-28-86291176, E-mail: jiary@sicau.edu.cn
About author:YANG Cui, E-mail: yc_itzel@163.com;
Supported by:
This study was supported by the National Science and Technology Support Program (2015BAD12B05), the China Agricultural Research System (CARS-43-8), the Integration and Demonstration of Key Technologies for Duck Industrial in Sichuan Province, China (2014NZ0030), the Ministry of Education Program of China (20125103110013), the Sichuan Province Research Programs, China (2013HH0042/2013TD0015/2014-002).

摘要/Abstract

Abstract: The capsid (Cap) protein, which is the only structural protein of duck circovirus (DuCV), is the most important antigen for the development of vaccines against DuCV and the virus’s serological diagnostic methods. In order to use yeast expression system to produce a large quantities of DuCV Cap protein which is close to its natural form to display the antigen peptides perfectly, the Cap gene was optimized into the codon-optimized capsid (Opt-Cap) gene towards the preference of yeast firstly. Then, the genes of Cap and Opt-Cap were separately cloned into pPIC9K plasmid and transformed into Picha pastoris GS115. The strains that displayed the phenotype of Mut+ and contained multiple inserts of expression cassette were selected from those colonies. After the induction expression, the secretory type of Cap protein, which was about 43 kDa, was best expressed under 0.5% (v/v) methanol and sorbitol induction. Compared with the Cap gene, the expression level of Opt-Cap gene was much higher. What’s more, the purified Cap protein had a good reactivity to its specific polyclone antibody and DuCV-positive serum, and it was able to self-assemble into virus-like particles (VLPs). These VLPs, with a diameter of 15–20 nm and without a nucleic acid structure, showed a high level of similarity to DuCV particles in size and shape. All of the results demonstrated that, based on the codon-optimization, it is suitable to use the P. pastoris expression system to produce DuCV VLPs on a large scale. It is the first time that a large amounts of DuCV VLPs were produced successfully in P. pastoris, which might be particularly useful for the further studies of serological diagnosis and vaccines of DuCV.

Key words: capsid gene , codon-optimization , duck circovirus , virus-like particles

. [J]. Journal of Integrative Agriculture, 2017, 16(07): 1601-1608.

YANG Cui, XU Yu, JIA Ren-yong, LIU Si-yang, WANG Ming-shu, ZHU De-kang, CHEN Shun, LIU Ma-feng, ZHAO Xin-xin, SUN Kun-feng, JING Bo, YIN Zhong-qiong, CHENG An-chun . The codon-optimized capsid gene of duck circovirus can be highly expressed in yeast and self-assemble into virus-like particles[J]. Journal of Integrative Agriculture, 2017, 16(07): 1601-1608.

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The codon-optimized capsid gene of duck circovirus can be highly expressed in yeast and self-assemble into virus-like particles

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