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首页> 外文期刊>Transgenic research >An optimized sericin-1 expression system for mass-producing recombinant proteins in the middle silk glands of transgenic silkworms
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An optimized sericin-1 expression system for mass-producing recombinant proteins in the middle silk glands of transgenic silkworms

机译：优化的丝胶蛋白1表达系统，用于在转基因蚕中部丝腺中大量生产重组蛋白

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摘要

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.
机译：蚕的中间丝腺（MSG）被认为是大量生产有价值的重组蛋白的潜在宿主。已经建立了基于丝胶蛋白1基因启动子的转基因MSG表达系统（sericin-1表达系统），以在MSG中产生各种重组蛋白。然而，仍然需要进一步修饰丝胶蛋白1表达系统的活性以产生更高量的重组蛋白。在这项研究中，我们提供了另一种修饰策略，可通过使用重庆市家蚕核多角体病毒（BmNPV）株和纤维蛋白的3'UTRs的hr3增强子（hr3 CQ）来构建高效的sericin-1表达系统。重链（Fib-HPA），纤维蛋白轻链（Fib-LPA）和Sericin1（Ser1PA）基因。我们首先分析了这些DNA元素对萤光素酶表达的影响，发现hr3 CQ和Ser1PA的组合最有效地增加了萤光素酶的活性。然后，hr3 CQ和Ser1PA被用来修饰sericin1表达系统。通过piggyBac转座子介导的遗传转化方法产生了带有这些修饰的sericin1表达载体的转基因蚕。我们的结果表明，含有hr3 CQ和Ser1PA的转基因蚕中DsRed报告基因的mRNA水平显着提高了9倍，达到内源性sericin1的83％。结果，重组RFP的产量增加了16倍，达到茧壳重量的9.5％（w / w）。我们得出的结论是，这种修饰的丝胶蛋白1表达系统是有效的，并且将有助于MSG作为宿主，大量生产有价值的重组蛋白。

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