Expression and characterization of recombinant human VEGF165 in the middle silk gland of transgenic silkworms
Expression and characterization of recombinant human VEGF165 in the middle silk gland of transgenic silkworms
复制标题
重组人VEGF165在转基因蚕中部丝腺中的表达及特性
DOI：
10.1007/s11248-019-00173-y
复制
发表时间：
2019
影响因子：
3
通讯作者：
Xu Hanfu
中科院分区：
文献类型：
--
作者：
Zhang Tianyang;Liu Rongpeng;Luo Qin;Qu Dawei;Chen Tao;Yao Ou;Xu Hanfu
重组人血管内皮生长因子 (rhVEGF) 在治疗性血管生成和抑制 VEGF 介导的病理性血管生成方面具有重要应用。此前的研究表明，rhVEGF可以在多种表达系统中产生，包括大肠杆菌、酵母、昆虫细胞和哺乳动物细胞。然而，人们对这种蛋白质在活体器官中的有效产生知之甚少。在这里，我们首次报道了rhVEGF165在转基因蚕系S1-V165的中丝腺（MSG）中的表达和特征。我们的结果证实：（1）rhVEGF165在MSG细胞中高表达，并分泌到S1-V165的茧中； (2)rhVEGF165的二聚体可以很容易地用碱性溶液从S1-V165茧中溶解； (3)从S1-V165茧中提取的rhVEGF165在培养的人脐静脉内皮细胞中表现出比hVEGF165标准品稍好的细胞增殖活性。本研究为利用转基因蚕的味精生产具有生物活性的 rhVEGF165 提供了另一种策略。
Recombinant human vascular endothelial growth factor (rhVEGF) has important applications in therapeutic angiogenesis and inhibition of VEGF-mediated pathological angiogenesis. Previous studies have shown that rhVEGF can be produced in several expression systems, includingEscherichia coli, yeasts, insect cells and mammalian cells. However, little is known regarding the effective production of this protein in organs of live organisms. Here, we report for the first time the expression and characterization of rhVEGF165 in the middle silk gland (MSG) of the transgenic silkworm line S1-V165. Our results confirmed that (1) rhVEGF165 was highly expressed in MSG cells and was secreted into the cocoon of S1-V165; (2) the dimeric form of rhVEGF165 could be easily dissolved from S1-V165 cocoons using an alkaline solution; (3) rhVEGF165 extracted from S1-V165 cocoons exhibited slightly better cell proliferative activity than the hVEGF165 standard in cultured human umbilical vein endothelial cells. This study provides an alternative strategy for the production of bioactive rhVEGF165 using the MSG of transgenic silkworms.