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The cytosolic Ca<jats:sup>2+<\/jats:sup> concentration is tightly regulated at the physiological cytosolic concentration of 50\u2013100\u00a0n<jats:sc>m<\/jats:sc>. The <jats:italic>Toxoplasma gondii<\/jats:italic> genome predicts the presence of several genes encoding potential Ca<jats:sup>2+<\/jats:sup> channels, pumps, and transporters. Many of these genes are weakly expressed and likely tightly regulated due to their potential impact to the physiology of the cell. Endogenous tagging has been widely used to localize proteins in <jats:italic>T.\u00a0gondii<\/jats:italic> but low level of expression of many of them makes visualization of tags difficult and sometimes impossible. The use of high\u2010performance tags for labeling proteins expressed at low level is ideal for investigating the localization of these gene products. We designed a Carboxy\u2010terminus tagging plasmid containing the previously characterized \u201cspaghetti monster\u2010<jats:styled-content style=\"fixed-case\">HA<\/jats:styled-content>\u201d (sm<jats:styled-content style=\"fixed-case\">HA<\/jats:styled-content>) or \u201cspaghetti monster\u2010<jats:styled-content style=\"fixed-case\">MYC<\/jats:styled-content>\u201d (sm<jats:styled-content style=\"fixed-case\">MYC<\/jats:styled-content>) tags. These tags consist of 10 copies of a single epitope (<jats:styled-content style=\"fixed-case\">HA<\/jats:styled-content> or <jats:styled-content style=\"fixed-case\">MYC<\/jats:styled-content>) inserted into a darkened green fluorescence protein scaffold. We localized six proteins of various levels of expression. Clonal lines were isolated and validated by <jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content>, western blot, and immunofluorescence analyses. Some gene products were only visible when tagged with sm<jats:styled-content style=\"fixed-case\">HA<\/jats:styled-content> and in one case the sm<jats:styled-content style=\"fixed-case\">HA<\/jats:styled-content> revealed a novel localization previously undetected.<\/jats:p>","DOI":"10.1111\/jeu.12626","type":"journal-article","created":{"date-parts":[[2018,4,19]],"date-time":"2018-04-19T19:25:56Z","timestamp":1524165956000},"page":"709-721","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":45,"title":["Tagging of Weakly Expressed <i>Toxoplasma gondii<\/i> Calcium\u2010Related Genes with High\u2010Affinity Tags"],"prefix":"10.1111","volume":"65","author":[{"given":"Miryam A.","family":"Hortua Triana","sequence":"first","affiliation":[{"name":"Center for Tropical and Emerging Global Diseases University of Georgia Athens Georgia 30602"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karla M.","family":"M\u00e1rquez\u2010Nogueras","sequence":"additional","affiliation":[{"name":"Center for Tropical and Emerging Global Diseases University of Georgia Athens Georgia 30602"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Le","family":"Chang","sequence":"additional","affiliation":[{"name":"Center for Tropical and Emerging Global Diseases University of Georgia Athens Georgia 30602"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew J.","family":"Stasic","sequence":"additional","affiliation":[{"name":"Center for Tropical and Emerging Global Diseases University of Georgia Athens Georgia 30602"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Catherine","family":"Li","sequence":"additional","affiliation":[{"name":"Center for Tropical and Emerging Global Diseases University of Georgia Athens Georgia 30602"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kevin A.","family":"Spiegel","sequence":"additional","affiliation":[{"name":"Center for Tropical and Emerging Global Diseases University of Georgia Athens Georgia 30602"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amrita","family":"Sharma","sequence":"additional","affiliation":[{"name":"Center for Tropical and Emerging Global Diseases University of Georgia Athens Georgia 30602"},{"name":"Department of Cellular Biology University of Georgia Athens Georgia 30602"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhu\u2010Hong","family":"Li","sequence":"additional","affiliation":[{"name":"Center for Tropical and Emerging Global Diseases University of Georgia Athens Georgia 30602"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2041-6295","authenticated-orcid":false,"given":"Silvia N.J.","family":"Moreno","sequence":"additional","affiliation":[{"name":"Center for Tropical and Emerging Global Diseases University of Georgia Athens Georgia 30602"},{"name":"Department of Cellular Biology University of Georgia Athens Georgia 30602"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2018,5,11]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/26.2.623"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tips.2015.09.001"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2133463100"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1018217108"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/312315a0"},{"key":"e_1_2_6_7_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M115.652511"},{"key":"e_1_2_6_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2016.02.006"},{"key":"e_1_2_6_9_1","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1042\/bj3420379","article-title":"Ethanol and acetaldehyde elevate intracellular [Ca2\u00a0+\u00a0] and stimulate microneme discharge in Toxoplasma gondii","volume":"342","author":"Carruthers V. 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