Novel tag protein for protein enrichment expression and intracellular localization as well as application thereof
A technology for labeling proteins and proteins, applied in the field of labeling proteins, can solve the problems of reducing cell activity, limiting protein yield, increasing production costs, etc., to achieve the effects of increasing final output, maintaining activity, and reducing loss
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[0059] Example 1. Expression of pEGFP-C1 and pEGFP-C1-RA1 in 293T cells
[0060] figure 1 .Confocal microscopy observation results of 293T cells transfected with pEGFP-C1 for 24 hours.
[0061] A) Excitation wavelength 488, signal collection channel 507, eGFP green fluorescence signal;
[0062] B) Excitation wavelength 364, signal collection channel 461, DAPI blue fluorescence signal;
[0063] C) Experimental results after the fusion of eGFP and DAPI signal channels;
[0064] D) Partially enlarged 5 times.
[0065] From figure 1 It can be seen from A that there is a large amount of green fluorescent signal (derived from the expression of eGFP) in the 293T cells successfully transfected with the pEGFP-C1 plasmid. figure 1 The blue signal in B is derived from DAPI. Due to the use of a membrane breaker (TritonX-100), no matter whether the cells are successfully transfected and express eGFP, the nucleus can be stained by DAPI, so the number of cells is greater than figure 1 A. will figure 1...
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[0073] Example 2. The expression of pEGFP-C1 and pEGFP-C1-RA1 in Hela cells
[0074] From image 3 It can be seen from A that there is a large amount of green fluorescent signal (derived from the expression of eGFP) in the Hela cells successfully transfected with the pEGFP-C1 plasmid. image 3 The blue signal in B is derived from DAPI. Due to the use of a membrane breaker (TritonX-100), no matter whether the cells are successfully transfected and express eGFP, the nucleus can be stained by DAPI, so the number of cells is greater than image 3 A. will image 3 After the images of A and 3B are fused, it can be found that the cells emitting eGFP green fluorescence overlap with part of the nuclei stained by DAPI. After local magnification (5 times) of the overlapping part, it can be found that the green fluorescence signal in the Hela cells successfully transfected with the pEGFP-C1 plasmid is evenly distributed, and the DAPI signal in the nucleus is blocked to a certain extent.
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