title: An optimized sericin-1 expression system for mass-producing recombinant proteins in the middle silk glands of transgenic silkworms.
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.
authors: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, 400715, People's Republic of China.
authors: Feng
authors: F
authors: Wang
authors: Hanfu
authors: H
authors: Xu
authors: Lin
authors: L
authors: Yuan
authors: Sanyuan
authors: S
authors: Ma
authors: Yuancheng
authors: Y
authors: Wang
authors: Xiaoli
authors: X
authors: Duan
authors: Jianping
authors: J
authors: Duan
authors: Zhonghuai
authors: Z
authors: Xiang
authors: Qingyou
authors: Q
authors: Xia
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics* / Q000502:physiology; D001709:Biotechnology / Q000379:methods*; D012831:Bombyx / Q000235:genetics* / Q000378:metabolism / Q000821:virology*; D002460:Cell Line; D004742:Enhancer Elements, Genetic / Q000235:genetics; D005349:Fibroins / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D008156:Luciferases / Q000378:metabolism; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D009841:Oligonucleotides / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D047030:Sericins / Q000235:genetics / Q000378:metabolism*; D018411:Spodoptera
chemical_list: D009841:Oligonucleotides; D011994:Recombinant Proteins; D047030:Sericins; C482288:fibroin, silkworm; D005349:Fibroins; D008156:Luciferases