Feng Yun, Yang Yunxia, Huang Ning, Wu Qi, Wang Boyao
Research Unit of Infection & Immunity, West China Medical Center of Sichuan University, Chengdu 610041.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2003 Dec;20(4):634-7.
This study was aimed at constructing a prokaryotic expression system to resolve the difficulties in acquiring antibacterial peptide and to meet the needs of research and drug development. Total RNA was extracted from human pulmonary gland epithelial cell line SPC-A-1, and a cDNA encoding mature FALL-39 peptide was amplified by RT-PCR. The recombinant prokaryotic expression vector pGEX-1 lambda T-FALL-39 was constructed. Using affinity chromatography, thrombin cleaving and AU-PAGE elution, we obtained the purified FALL-39. MIC, MEC, MBC analyses demonstrated that the FALL-39 had strong antibacterial activity.
本研究旨在构建原核表达系统，以解决抗菌肽获取困难的问题，并满足研究和药物开发的需求。从人肺腺上皮细胞系SPC-A-1中提取总RNA，通过RT-PCR扩增编码成熟FALL-39肽的cDNA。构建重组原核表达载体pGEX-1λT-FALL-39。通过亲和层析、凝血酶切割和AU-PAGE洗脱，我们获得了纯化的FALL-39。MIC、MEC、MBC分析表明，FALL-39具有较强的抗菌活性。
