Qian Qiujie, You Zhengying, Ye Lupeng, Che Jiaqian, Wang Yiran, Wang Shaohua, Zhong Boxiong
College of Animal Sciences, Zhejiang University, Hangzhou, P. R. China.
Haining Sericulture Technology Extension Station of Zhejiang Province, Haining Zhejiang, China.
PLoS One. 2018 Jan 19;13(1):e0191507. doi: 10.1371/journal.pone.0191507. eCollection 2018.
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.
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。在本研究中,将源自pESNT-PBase质粒的融合转座酶的mRNA与一个pBHSA质粒共同注射到受精卵中,该pBHSA质粒含有在丝素轻链(FL)启动子控制下的HSA基因。成功获得了56个在后丝腺(PSG)中表达外源重组HSA(rHSA)且具有稳定遗传的转基因家蚕品系。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盐水(PBS)提取。在我们的研究中,分离得到的最高量的rHSA占茧壳总可溶性蛋白的29.1%,这表明转基因家蚕在茧壳中平均产生17.4μg/mg的rHSA。通过培育转基因家蚕在PSG中生产可溶性rHSA是一种新方法,借此可以通过简单饲养家蚕大量生产无病毒且有功能的HSA。