NDLI: An optimized sericin-1 expression system for mass-producing recombinant proteins in the middle silk glands of transgenic silkworms

Content Provider	Springer Nature Link
Author	Wang, Feng Xu, Hanfu Yuan, Lin Ma, Sanyuan Wang, Yuancheng Duan, Xiaoli Duan, Jianping Xiang, Zhonghuai Xia, Qingyou
Copyright Year	2013
Abstract	The middle silk gland (MSG) of silkworm is thought to be a potential host for mass-producing valuable recombinant proteins. Transgenic MSG expression systems based on the usage of promoter of sericin1 gene (sericin-1 expression system) have been established to produce various recombinant proteins in MSG. However, further modifying the activity of the sericin-1 expression system to yield higher amounts of recombinant proteins is still necessary. In this study, we provide an alternative modification strategy to construct an efficient sericin-1 expression system by using the hr3 enhancer (hr3 CQ) from a Chongqing strain of the Bombyx mori nuclear polyhedrosis virus (BmNPV) and the 3′UTRs of the fibroin heavy chain (Fib-HPA), the fibroin light chain (Fib-LPA), and Sericin1 (Ser1PA) genes. We first analyzed the effects of these DNA elements on expression of luciferase, and found that the combination of hr3 CQ and Ser1PA was most effective to increase the activity of luciferase. Then, hr3 CQ and Ser1PA were used to modify the sericin1 expression system. Transgenic silkworms bearing these modified sericin1 expression vectors were generated by a piggyBac transposon mediated genetic transformation method. Our results showed that mRNA level of DsRed reporter gene in transgenic silkworms containing hr3 CQ and Ser1PA significantly increased by 9 fold to approximately 83 % of that of endogenous sericin1. As the results of that, the production of recombinant RFP increased by 16 fold to 9.5 % (w/w) of cocoon shell weight. We conclude that this modified sericin-1 expression system is efficient and will contribute to the MSG as host to mass produce valuable recombinant proteins.
Starting Page	925
Ending Page	938
Page Count	14
File Format	PDF
ISSN	09628819
Journal	Transgenic Research
Volume Number	22
Issue Number	5
e-ISSN	15739368
Language	English
Publisher	Springer Netherlands
Publisher Date	2013-02-23
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Transgene Sericin-1 expression system Modification Silkworm Molecular Medicine Plant Genetics & Genomics Animal Genetics and Genomics Plant Sciences Human Genetics
Content Type	Text
Resource Type	Article
Subject	Genetics Animal Science and Zoology Biotechnology Agronomy and Crop Science

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