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Production of human type II collagen using an efficient baculovirus-silkworm multigene expression system

文献类型： 外文期刊

作者： Qi, Qi 1 ; Yao, Lunguang 1 ; Liang, Zhisheng 3 ; Yan, Donghua 1 ; Li, Zhuo 4 ; Huang, Yadong 3 ; Sun, Jingchen 1 ;

作者机构： 1.South China Agr Univ, Coll Anim Sci, Subtrop Sericulture & Mulberry Resources Protect, Guangdong Prov Key Lab Agroanim Genom & Mol Breed, Guangzhou 510642, Guangdong, Peoples R China

2.Nan Yang Normal Univ, Henan Key Lab Ecol Secur Water Source Reg Midline, Nanyang 473000, Henan, Peoples R China

3.Jinan Univ, Inst Biomed, Guangzhou 510632, Guangdong, Peoples R China

4.Nanyang Med Univ, Coll Med Techno

关键词： Human type II collagen;BEVS;Bombyx mori;Sf9;Triple-helix

期刊名称：MOLECULAR GENETICS AND GENOMICS （ 影响因子：3.291； 五年影响因子：3.257 ）

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收录情况： SCI

摘要： Human type II collagen is a macromolecular protein found throughout the human body. The baculovirus expression vector system is one of the most ideal systems for the routine production and display of recombinant eukaryotic proteins in insect, larvae, and mammalian cells. We use this system to express a full-length gene, human type II collagen cDNA (4257 bp), in cultured Spodoptera frugiperda 9 cells (Sf9), Bombyx mori cells, and silkworm larvae. In this study, the expression of human type II collagen gene in both insect cells and silkworm larvae was purified by nickel column chromatography, leading to 300-kDa bands in SDS-PAGE and western blotting indicative of collagen alpha-chains organized in a triple-helical structure. About 1 mg/larva human type II collagen is purified from silkworm skin, which shows a putative large scale of collagen production way. An activity assay of recombinant human type II collagen expressed by silkworm larvae demonstrated that the recombinant protein has considerable bioactive properties. Scanning electron microscopy of purified proteins clearly reveals randomly distributed and pitted structures. We conclude that the baculovirus-silkworm multigene expression system can be used as an efficient platform for express active human type II collagen and other complicated eukaryotic proteins.

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