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. 2021 Feb 4;20:625–634. doi: 10.1016/j.omtm.2021.01.014

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© 2021 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Screening for positive colonies for the IL-5-expressing virus

(A) After transformation of GA products, eight colonies were chosen (lanes 2–9) and a colony PCR was conducted. Primers flanking the CMV and IL-15 coding region were used. As a positive control, the GA of CMV-IL-5-poly(A) (lane 10) was used. Lane 1 represents a 1 kb gene ruler from Thermo Fisher. (B) Simulation of expected lanes where unmodified adenovector is cut with EcoRI (lane 3) compared to positive adenovirus clones successfully having the gp19k replaced with the GOI (lane 2). Lane1 represents a 1 kb gene ruler from Thermo Fisher. (B) Actual representation of three colonies transformed with the assembled adenovirus containing the GOI (lanes 2, 3, and 4). Lane 5 represents wild-type adenovirus cut with EcoRI, while lane 1 represents a 1 kb gene ruler from Thermo Fisher. (C) Three samples of Sanger sequencing from one of the positive sequences. Original sequence represents the reference sequence of IL-15.