Liu Bainan, Kong Qianqian, Zhang Dong, Yan Lingli
Department of Immunology, Zunyi Medical University, Zunyi, 563000 Guizhou China.
3 Biotech. 2018 Apr;8(4):210. doi: 10.1007/s13205-018-1234-y. Epub 2018 Apr 6.
37-kDa immature laminin receptor protein (iLRP), the precursor of 67-kDa laminin receptor protein (LRP), is overexpressed on the surface of most cancer cells and recognized as a universal tumor antigen. The role makes it a potential target for cancer immunotherapy, which has been well-studied. Our study aimed to produce high quality of human iLRP in bacteria so that the needs in research of its clinical application could be met. The powerful system for heterologous protein expression, pET system was used. Two types of DNA sequences encoding the same amino acid sequences were separately cloned into the vector pET30a(+). One of the resulting vectors includes the wild-type , and other one includes the codon-optimized . The expression by both genes was then compared in BL21(DE3). Our results revealed that the performance of codon optimization was crucial for the expression of human in . The yield was significantly enhanced up to 300 mg/L of bacterial culture by this approach.
37 kDa未成熟层粘连蛋白受体蛋白（iLRP）是67 kDa层粘连蛋白受体蛋白（LRP）的前体，在大多数癌细胞表面过度表达，被认为是一种通用肿瘤抗原。这一特性使其成为癌症免疫治疗的潜在靶点，对此已有深入研究。我们的研究旨在利用细菌高效生产人iLRP，以满足其临床应用研究的需求。我们采用了强大的异源蛋白表达系统——pET系统。将编码相同氨基酸序列的两种DNA序列分别克隆到载体pET30a(+)中。其中一个构建载体包含野生型序列，另一个包含密码子优化后的序列。然后在BL21(DE3)中比较这两个基因的表达情况。我们的结果表明，密码子优化对人iLRP在大肠杆菌中的表达至关重要。通过这种方法，细菌培养物中的产量显著提高，达到了300 mg/L。
