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PMC:7795856 / 7542-9075 JSONTXT

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LitCovid-PubTator

Id Subject Object Predicate Lexical cue tao:has_database_id
175 0-3 Gene denotes 2.1 Gene:6700
183 115-118 Gene denotes Tα1 Gene:134864
184 223-226 Gene denotes Tα1 Gene:134864
185 517-520 Gene denotes Tα1 Gene:134864
186 217-222 Species denotes human Tax:9606
187 349-356 Species denotes E. coli Tax:562
188 725-732 Species denotes E. coli Tax:562
189 793-796 Chemical denotes tet MESH:C010349
195 1173-1176 Gene denotes Tα1 Gene:134864
196 1279-1301 Chemical denotes sodium dodecyl sulfate MESH:D012967
197 1302-1316 Chemical denotes polyacrylamide MESH:C016679
198 1338-1341 Chemical denotes SDS MESH:D012967
199 1437-1440 Chemical denotes SDS MESH:D012967

LitCovid-sentences

Id Subject Object Predicate Lexical cue
T48 0-4 Sentence denotes 2.1.
T49 5-54 Sentence denotes Cloning and Bacterial Production of PASylated Tα1
T50 55-448 Sentence denotes To achieve C-terminal PASylation of N-terminally acetylated Tα1, a plasmid harboring a bicistronic operon was constructed to allow the simultaneous expression of human Tα1 (UniProtKB ID: P06454; residues 2–29), C-terminally fused with a PAS polypeptide comprising 601 amino acids [34], and the E. coli N-acetyltransferase RimJ (UniProtKB ID: P0A948) based on the vector pASK75 (Figure 1) [36].
T51 449-631 Sentence denotes In a parallel attempt, a plasmid encoding an N-terminally PASylated Tα1 was constructed, again, using plasmid pASK75 as the backbone (this time omitting the RimJ cistron, see below).
T52 632-820 Sentence denotes Cytoplasmic gene expression was performed in both cases on a 2 L shake flask scale using the E. coli strain NEBexpress under control of the chemically inducible tet promoter/operator [36].
T53 821-910 Sentence denotes The whole cell lysate was analyzed prior to and 15 h after induction by Western blotting.
T54 911-1216 Sentence denotes Using a monoclonal antibody that recognizes an epitope of the PAS#1 sequence, a distinct band with an approximate molecular size above 250 kDa was detected (Figure 1b), which demonstrated successful bacterial expression of the full-length C-terminally PASylated Tα1 (Tα1-PAS) without signs of degradation.
T55 1217-1533 Sentence denotes Of note, the unusually slow migration of Tα1-PAS (52.7 kD) in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) is well known for PASylated proteins [31,37] and can be explained by the poor binding of SDS (which provides the electrophoretic driving force) to the strongly hydrophilic PAS sequence.