结核分枝杆菌荧光标记菌株

荧光蛋白可以用于标记蛋白，在特定激发光激发下会发出特定的荧光，广泛应用于活细胞实时观察。荧光蛋白标记结核分枝杆菌特定蛋白后，可以研究该蛋白的定位，测定结核分枝杆菌氧化还原水平，研究结核分枝杆菌在动物细胞中的动态变化等，有助于结核分枝杆菌免疫学领域的研究。

Figure 7. M.tb localizes to LC3-positive compartments. (A) GFPLC3expressing macrophages were infected with mCherry-M.tb (MOI 50

or 10, 6 h) and then fixed and observed by confocal microscopy. At high MOI there are GFP-LC3 rings surrounding (arrowhead) around mCherry-M.tb and co-localization indicated by yellow pixels (arrow) that are not seen in macrophages challenged at low MOI. (B) LAMP1 immunostaining of GFP-LC3 cell line macrophages infected with M.tb Edrman (MOI 50 or 10, 5 h) demonstrates co-localization of LC3 and lysosomal markers (arrowheads).

doi:10.1371/journal.pone.0027972.g007

Figure 2. Differences in expression of rv2390c9::GFP in Mtb present in vaccinated versus mock-treated mice. Erdman(rv2390c9::GFP, smyc9::mCherry) was inoculated into vaccinated or mock-treated C57BL/6J WT mice for up to 56 days. (A) shows 3D confocal images from a 14, 28, or 42 day infection. All bacteria are marked in red (smyc9::mCherry), reporter signal is shown in green (rv2390c9::GFP), nuclei are marked in grayscale (DAPI), and phalloidin staining of f-actin is shown in blue. Scale bar 10 mm. (B) shows quantification of the GFP/mm3 signal for each bacterium measured from multiple 3D confocal images, at the indicated time points. Each point on the graph represents a bacterium or a tightly clustered group of bacteria (mock-treated – filled symbols, vaccinated – open symbols). Horizontal lines mark the median value for each group. p-values were obtained with a Mann-Whitney statistical test.

doi:10.1371/journal.ppat.1004394.g002

应用分枝杆菌过表达质粒pMV261（游离型）、pMV361（整合型），构建eGFP（绿色荧光）、mCherry（红色荧光）荧光质粒，使荧光蛋白在分枝杆菌内持续表达，感染巨噬细胞或其他细胞，检测感染效率、细胞自噬、细胞凋亡、结核分枝杆菌存活能力等作用；感染小鼠，检测结核分枝杆菌定位；另外，荧光菌株还可用于肺外结核如肠结核、脑结核、骨结核等定位研究，助力于结核分枝杆菌致病机制的研究。

荧光菌株示例图

注：图A为耻垢分枝杆菌 eGFP、mCherry荧光菌株在蓝光激发下发出绿色和红色荧光；图B为牛结核分枝杆菌BCG eGFP、mCherry荧光菌株在蓝光激发下发出绿色和红色荧光；图C为结核分枝杆菌H37Ra eGFP、mCherry荧光菌株在蓝光激发下发出绿色和红色荧光

晶诺生物已有耻垢分枝杆菌、牛结核分枝杆菌BCG、结核分枝杆菌H37Rv、结核分枝杆菌H37Ra的红色荧光（mCherry）、绿色荧光（eGFP）菌株，欢迎各位老师订购。正在研发蓝色荧光（BFP）、黄色荧光（YFP）、近红外（iRFP720）、荧光素酶等标记的分枝杆菌菌株，请各位老师持续关注！

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