Method for marking acinetobacter baumannii by green fluorescent protein gene
A technology of Acinetobacter baumannii and green fluorescent protein, applied in the biological field, can solve the problems of life-threatening patients
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Embodiment 1A
[0022] Example 1Ab-GFP strain construction
[0023] 1. Experimental materials
[0024] Human alveolar epithelial cells (HPAEpiC) cell line was purchased from CELLBIO (No.: CBR130583), RPMI 1640 (Gino, GNM31800), RPMI 1640 double antibody (Gino, GNM-31800-S), PBS buffer 1× (Gino, GNM20012), 0.25% trypsin + EDTA (Gino, GNM-25260), fetal bovine serum (CAPRICORNSCIENTIFIC, FBS-11A), agar powder (Best, BS1068), peptone (Best, BS7006 ), yeast powder (Best, BS1069), sodium chloride (Sinopharm Chemical Reagent Co., Ltd.), Triton X-100 (polyethylene glycol octylphenyl ether, Dalian Meilun Biotechnology Co., Ltd., MB2486), iFluor TM 647phalloidin (phalloidin-iFluor647 conjugate, Dalian Meilun Biotechnology Co., Ltd., MB5939), DAPI (4', 6-diamidino-2-phenylindole, Thermo, 62247), paraformaldehyde ( McLean, P804536-500g), bovine serum albumin (Shanghai Yuanye Biotechnology Co., Ltd., S25762), 10× polylysine (Dalian Meilun Biotechnology Co., Ltd., MA0174). The strains, plasmids and pri...
Embodiment 2
[0063] 1. Construction of fluorescently labeled Acinetobacter baumannii infection cell model
[0064] Acinetobacter baumannii mainly infects the respiratory tract and causes the occurrence and development of related respiratory infection diseases. Therefore, in the present invention, human alveolar epithelial cells (Human alveolar epithelial cells, HPAEpiC, cell strain purchased from CELLBIO, number: CBR130583) are used as A cellular model for studying bacteria-host interactions. The result is as Figure 4 As shown, by fluorescence microscope observation, the model of infection of host cells by the GFP-labeled strain of Acinetobacter baumannii can be successfully constructed, and the relative position, adhesion or invasion of bacteria and host cells can be accurately located. Provide basic experimental techniques for dynamic real-time observation of bacterial-cell interaction research, as well as biological functions of Acinetobacter baumannii-related pathogenic factors.
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