克服问题以生产. 的重组蛋白 LipL21。
Overcoming problems to produce the recombinant protein LipL21 of .
机构信息
Programa de Pós-Graduação Interunidades em Biotecnologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, Avenida Lineu Prestes, São Paulo, 2415 - 05508-900, Brazil.
Laboratório de Desenvolvimento de Vacinas, Instituto Butantan, Avenida Vital Brazil, São Paulo, 1500 - 05503-900, Brazil.
出版信息
Biotechniques. 2023 Mar;74(3):137-142. doi: 10.2144/btn-2022-0076. Epub 2023 Apr 17.
The production of leptospiral recombinant proteins in the soluble form and in high yield from is still a challenge. This work presents the cloning, expression and purification of the outer membrane protein of , LipL21, which is considered an interesting target for vaccine and diagnostics development. The expression profile and yield of LipL21 was compared after cloning in the vectors pAE, pET28a and pET-SUMO, and it was observed that LipL21 was expressed in a low amount with pAE vector. By using the pET-28a vector, protein expression was increased, but the majority of the product was obtained as inclusion bodies. As a highlight, using a pET-SUMO vector was shown to overcome the problems of low expression and solubility of the lipoprotein LipL21.
从 中以可溶形式和高产率生产钩端螺旋体重组蛋白仍然是一个挑战。本工作展示了 的外膜蛋白 LipL21 的克隆、表达和纯化，LipL21 被认为是疫苗和诊断开发的一个有趣的靶标。在 pAE、pET28a 和 pET-SUMO 载体中克隆后，比较了 LipL21 的表达谱和产量，结果表明 pAE 载体表达的 LipL21 量较低。使用 pET-28a 载体可以增加蛋白表达，但大部分产物是以包涵体的形式获得的。值得一提的是，使用 pET-SUMO 载体可以克服脂蛋白 LipL21 表达量低和可溶性差的问题。
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