LitCovid-sentences  

PMC:7019868 / 7501-9327 JSONTXT 7 Projects

Annnotations TAB TSV DIC JSON TextAE

Id Subject Object Predicate Lexical cue
T54 0-76 Sentence denotes The strategy used to recover iPEDVPT-P96 has been described previously [19].
T55 77-186 Sentence denotes The approach to constructing a cDNA clone of iPEDVPT-P5 was technically identical to that of the iPEDVPT-P96.
T56 187-454 Sentence denotes However, we split the plasmid B into two fragments because the sequence remained toxic to the One Shot™ TOP10 Chemically Competent E. coli cells (Invitrogen, Carlsbad, USA) despite propagation in LB broth supplemented with 10% SOC medium and being incubated at 30 °C.
T57 455-653 Sentence denotes To generate chimeric viruses carrying heterologous spike (S) genes, namely the iPEDVPT-P5-96S and iPEDVPT-P96-5S, cDNA clones of iPEDVPT-P5 and iPEDVPT-P96, respectively, were used as the backbones.
T58 654-780 Sentence denotes The sequences covering the complete S gene of each virus were exchanged without disruption to the remaining genomic structure.
T59 781-881 Sentence denotes Sequence differences in the S genes of iPEDVPT-P5 and iPEDVPT-P96 viruses are summarized in Table 1.
T60 882-1074 Sentence denotes Each plasmid was digested with corresponding type-IIS restriction enzymes as designated in Figure S1, gel-purified, assembled and phenol-chloroform extracted to generate the full-length cDNAs.
T61 1075-1393 Sentence denotes The cDNAs were then in vitro transcribed to the full-length RNA transcripts using a mMessage mMachine T7 transcription kit (Ambion, Austin, CA, USA) and immediately electroporated into 800 μL of 107 cells/mL Vero cells in RNase-free phosphate buffered saline (PBS) along with 5 μg of PEDV nucleocapsid (N) transcripts.
T62 1394-1586 Sentence denotes After electroporation, the cells were allowed to recover in growth medium for approximately 16 h and then maintained in PI-medium until cytopathic effects involved over 90% of cell monolayers.
T63 1587-1721 Sentence denotes The whole flasks were subjected to one freeze-and-thaw cycle and the rescued viruses were passaged once to generate viral stocks (P1).
T64 1722-1826 Sentence denotes The viral stocks were titrated on Vero cells in 96-well plates to determine the viral titer (see below).