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High-efficiency production of human serum albumin in the posterior silk glands of transgenic silkworms Bombyx mori L

机译:在转基因蚕Bombyx mori L的后丝腺中高效生产人血清白蛋白

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摘要

Human serum albumin (HSA) is an important biological preparation with a variety of biological functions in clinical applications. In this study, the mRNA of a fusion transposase derived from the pESNT-PBase plasmid and a pBHSA plasmid containing the HSA gene under the control of a fibroin light chain (FL) promoter were co-injected into fertilized eggs. Fifty-six transgenic silkworm pedigrees expressing theexogenous recombinant HSA (rHSA) in the posterior silk glands (PSGs) with stable inheritance were successfully obtained. The SDS-PAGE and Western blot results confirmed that the rHSA was secreted into the transgenic silkworm cocoon, and the rHSA could be easily extracted with phosphate-buffered saline (PBS). In our research, the isolated highest amount rHSA constituted up to 29.1% of the total soluble protein of the cocoon shell, indicating that the transgenic silkworm produced an average of 17.4 μg/mg of rHSA in the cocoon shell. The production of soluble rHSA in the PSGs by means of generating transgenic silkworms is a novel approach, whereby a large amount of virus-free and functional HSA can be produced through the simple rearing of silkworms.
机译:人血清白蛋白(HSA)是一种重要的生物制剂,在临床应用中具有多种生物学功能。在这项研究中,将来自pESNT-PBase质粒的融合转座酶的mRNA和在纤维蛋白轻链(FL)启动子控制下的含有HSA基因的pBHSA质粒共注射到受精卵中。成功获得了56个在后部丝腺(PSGs)中表达稳定遗传的重组HSA(rHSA)的转基因家系。 SDS-PAGE和Western blot结果证实,rHSA被分泌到转基因蚕茧中,并且rHSA可以很容易地用磷酸盐缓冲液(PBS)提取。在我们的研究中,分离出的最高含量的rHSA占茧壳总可溶性蛋白的29.1%,这表明转基因蚕在茧壳中平均产生17.4μg/ mg的rHSA。通过产生转基因家蚕在PSG中生产可溶性rHSA是一种新颖的方法,通过简单饲养家蚕可以生产大量无病毒且功能正常的HSA。

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