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. 2021 Sep 22;13(10):1893. doi: 10.3390/v13101893

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Optimisation of expression construct (a) H1N1 NA constructs used in this study. Each horizonal bar represents one of the four constructs used in this study. The secretion signal (SS) is shown in yellow, the His-tag is shown in green, the tetramerisation domain (TET) is shown in purple, the thrombin cleavage site is shown in red. This image was made with IBS software [65]. (b) Relative neuraminidase activity, plotted following normalization to the sample with the highest activity: pTT5-NA (+SS−TET). (c–d) SDS-Page gels (4–20%) from expression trials of NA (+SS +TET), (−SS +TET), (+SS −TET), (−SS −TET). The supernatant samples were concentrated 10× before loading. Untransfected cells (untrans) represent a negative control for NA expression. (c) shows expression of NA from constructs that did not contain a tetramerisation domain (−TET). (d) shows expression of NA from constructs which contained a tetramerisation domain (+TET). Both gels were stained with Coomassie Blue.

Optimisation of expression construct (a) H1N1 NA constructs used in this study. Each horizonal bar represents one of the four constructs used in this study. The secretion signal (SS) is shown in yellow, the His-tag is shown in green, the tetramerisation domain (TET) is shown in purple, the thrombin cleavage site is shown in red. This image was made with IBS software [65]. (b) Relative neuraminidase activity, plotted following normalization to the sample with the highest activity: pTT5-NA (+SS−TET). (c–d) SDS-Page gels (4–20%) from expression trials of NA (+SS +TET), (−SS +TET), (+SS −TET), (−SS −TET). The supernatant samples were concentrated 10× before loading. Untransfected cells (untrans) represent a negative control for NA expression. (c) shows expression of NA from constructs that did not contain a tetramerisation domain (−TET). (d) shows expression of NA from constructs which contained a tetramerisation domain (+TET). Both gels were stained with Coomassie Blue.