Maidin Mohd Shawal Thakib, Song Adelene Ai-Lian, Jalilsood Tannaz, Sieo Chin Chin, Yusoff Khatijah, Rahim Raha Abdul
Department of Cell and Molecular Biology, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia; Institute of Bioscience Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
Plasmid. 2014 Jul;74:32-8. doi: 10.1016/j.plasmid.2014.05.003. Epub 2014 May 28.
A vector that drives the expression of the reporter gusA gene in both Lactobacillus plantarum and Lactococcus lactis was constructed in this study. This vector contained a newly characterized heat shock promoter (Phsp), amplified from an Enterococcus faecium plasmid, pAR6. Functionality and characterization of this promoter was initially performed by cloning Phsp into pNZ8008, a commercial lactococcal plasmid used for screening of putative promoters which utilizes gusA as a reporter. It was observed that Phsp was induced under heat, salinity and alkaline stresses or a combination of all three stresses. The newly characterized Phsp promoter was then used to construct a novel Lactobacillus vector, pAR1801 and its ability to express the gusA under stress-induced conditions was reproducible in both Lb. plantarum Pa21 and L. lactis M4 hosts.
本研究构建了一种能在植物乳杆菌和乳酸乳球菌中驱动报告基因gusA表达的载体。该载体包含一个新鉴定的热休克启动子（Phsp），它是从粪肠球菌质粒pAR6中扩增得到的。该启动子的功能和特性最初是通过将Phsp克隆到pNZ8008中来进行研究的，pNZ8008是一种用于筛选假定启动子的商业乳球菌质粒，它利用gusA作为报告基因。结果发现，Phsp在热、盐和碱性胁迫或这三种胁迫的组合下被诱导。然后，利用新鉴定的Phsp启动子构建了一种新型的植物乳杆菌载体pAR1801，其在胁迫诱导条件下表达gusA的能力在植物乳杆菌Pa21和乳酸乳球菌M4宿主中均可重现。
