蛛丝蛋白具有良好的生物相容性和生物可降解性，令其在生物医学材料如药物控释载体、手术缝合线、细胞培养支架、器官移植等方面的应用研究越来越广，但采用传统基因重组技术表达的重组蛛丝蛋白存在相对分子质量偏小、不具有分子取向等缺陷，使得用这种重组蛛丝蛋白纺制的丝纤维的力学性能不能令人满意。本研究利用大肠杆菌表达一种由蛛丝蛋白和类胶原蛋白融合构成的拟蛛丝蛋白，其为“ABA”型三嵌段共聚物，两端(A 段) 为(Pro-Gly-Pro)n 同聚体延伸段，可定向形成胶原三螺旋结构，中间(B 段) 为优化设计的蛛丝蛋白。利用三嵌段共聚物定向自组装形成超分子的特性，确保人工蛛丝纤维对相对分子质量的要求，也赋予拟蛛丝蛋白确定的分子取向，促进丝纤维的形成。本研究利用一对同尾酶，对拟蛛丝蛋白单体基因及类胶原蛋白基因进行同向重复串联，构建含拟蛛丝蛋白基因载体，最终在大肠杆菌中实现成功表达。
Abstract
Due to its good biocompatibility and biodegradability, spider silk proteins have found more and more applications in biomedical materials, such as drug controlled-release carriers, surgical sutures, cell culture stents, organ transplantation and so on. However, the recombinant spider silk protein expressed by traditional gene recombination technology has relatively small molecular weight and no molecular orientation. As a result, the mechanical properties of the silk fibers made with this recombinant spider silk protein are unsatisfactory. Here we intended to express a quasi spider silk protein composed of spider silk protein and collagen-like peptides using Escherichia coli. The quasi spider silk protein is a kind of “ABA-type” triblock copolymer. The triple helix-forming A blocks at the both ends of the triblock copolymers consist of (Pro-Gly-Pro)n homopolymeric stretches, and the middle section of the molecule (B section) consists of spider silk protein which has been designed optimally. The characteristics of the three block copolymer forming supramolecular by directed self-assembly ensure that artificial spider silk fibers will meet their requirement for relative molecular weight and certain molecular orientation to promote the formation of silk fibers. Having used a pair of isocaudarner, we succesfully constructed a quasi spider silk protein gene vector by monomer tandem repeating strategy carried out on spider silk protein monomer gene and collagen-like peptides gene, finally, the quasi spider silk protein gene had been expressed in Escherichia coli.
关键词
拟蛛丝蛋白 /
定向自组装 /
大肠杆菌 /
类胶原蛋白
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Key words
quasi spider silk protein /
directed self-assembly /
Escherichia coli /
collagen-like peptide
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