家蚕丝腺生物反应器表达狂犬病病毒核蛋白

doi:10.3864/j.issn.0578-1752.2013.18.023

Abstract

Abstract: 【Objective】In order to obtain a potent technology which can produce high value RVNP protein and then lay a foundation for the production of vaccines, the silk gland bioreactor was employed as an expression system in this study.【Method】The nucleoprotein gene of rabies virus ERA strain (RVNP) was cloned by RT-PCR, and the RVNP was firstly connected to the downstream of middle silk gland-specific Ser1 promoter, and then inserted into pBA3EGFP, an expression vector which contained an EGFP reporter gene. By using the transgenic method, the silk gland bioreactor of piggyback-mediated transgenic silkworm was obtained. 【Result】Twenty-six broods of transgenic silkworms (G1) were obtained and the GFP-positive percentage of broods was 59.09%. Further PCR identification indicated that RVNP had been integrated into silkworm genome, and Western blot analysis showed that the protein might be attached to the sericin which was secreted into the cocoon with the spinning of transgenic silkworms. 【Conclusion】Transgenic silkworm strain able to express exogenous RVNP protein was obtained. The present system is expected to become a high efficiency, low-cost and stable technical system to produce genetic engineering rabies vaccine.

Key words: rabies , silk gland bioreactor , piggyBac , silkworm (Bombyx mori)

YOU Zheng-Ying-1, WEI Hao-13, RUAN Xi-Zhen-2, YE Lu-Peng-1, WANG Shao-Hua-1, ZHOU Ji-Yong-2, ZHONG Bo-Xiong-1. Expression of RVNP in the Silk Gland of Transgenic Silkworm (Bombyx mori)[J].Scientia Agricultura Sinica, 2013, 46(18): 3922-3929.

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https://www.chinaagrisci.com/EN/Y2013/V46/I18/3922

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