High yield exogenous protein HPL production in the Bombyx mori silk gland provides novel insight into recombinant expression systems.
High yield exogenous protein HPL production in the Bombyx mori silk gland provides novel insight into recombinant expression systems.
复制标题
家蚕丝腺中高产外源蛋白 HPL 的产生为重组表达系统提供了新的见解。
DOI：
10.1038/srep13839
复制
发表时间：
2015-09-15
影响因子：
4.6
通讯作者：
Xu S
中科院分区：
文献类型：
--
作者：
Wang H;Wang L;Wang Y;Tao H;Yin W;SiMa Y;Wang Y;Xu S
家蚕丝腺（BmSG）以其上级蛋白质合成和分泌能力而受到广泛关注。然而，BmSG生物反应器的应用仍然是一个有争议的问题，因为重组蛋白的产量低。本研究设计了一个全长3057 bp的Hpl基因编码序列，并将其转化家蚕基因组，获得了Hpl在家蚕后部SG（BmPSG）中特异表达的突变体（Hpl/Hpl）。在突变体中，Fib-L和P25以及相应的编码蛋白的转录水平没有降低。但Fib-H mRNA水平下降了71.1%，分泌丝素中Fib-H蛋白从91.86%下降到71.01%。Hpl的mRNA水平分别为Fib-H和Fib-L的0.73%和0.74%，而HPL蛋白占丝素蛋白的18.85%和分泌丝蛋白总量的15.46%。因此，外源蛋白被非常有效地翻译和分泌。进一步对BmPSG细胞差异表达基因（DEG）进行分析，共检测到891个DEG，其中208个与蛋白质代谢相关。降低内源丝蛋白在BmPSG中的表达量可以有效提高外源重组蛋白的生产效率。
The silk gland of Bombyx mori (BmSG) has gained significant attention by dint of superior synthesis and secretion of proteins. However, the application of BmSG bioreactor is still a controversial issue because of low yields of recombinant proteins. Here, a 3057 bp full-length coding sequence of Hpl was designed and transformed into the silkworm genome, and then the mutant (Hpl/Hpl) with specific expression of Hpl in posterior BmSG (BmPSG) was obtained. In the mutants, the transcription level of Fib-L and P25, and corresponding encoding proteins, did not decrease. However, the mRNA level of Fib-H was reduced by 71.1%, and Fib-H protein in the secreted fibroin was decreased from 91.86% to 71.01%. The mRNA level of Hpl was 0.73% and 0.74% of Fib-H and Fib-L, respectively, while HPL protein accounted for 18.85% of fibroin and 15.46% of the total amount of secreted silk protein. The exogenous protein was therefore very efficiently translated and secreted. Further analysis of differentially expressed gene (DEG) was carried out in the BmPSG cells and 891 DEGs were detected, of which 208 genes were related to protein metabolism. Reduced expression of endogenous silk proteins in the BmPSG could effectively improve the production efficiency of recombinant exogenous proteins.