Matches in SemOpenAlex for { <https://semopenalex.org/work/W2374663397> ?p ?o ?g. }
Showing items 1 to 80 of
80
with 100 items per page.
- W2374663397 endingPage "7, 42" @default.
- W2374663397 startingPage "33" @default.
- W2374663397 abstract "To construct the eukaryotic expression system of L.interrogans lipL32/1-ompL1/1 fusion gene and to identify the immunoreactivity of expression products.PCR with linking primer was used to construct the fusion gene lipL32/1-ompL1/1. The P.pastoris eukaryotic expression system of the fusion gene, pPIC9K-lipL32/1-ompL1/1-P. pastorisGS115, was constructed after the fusion gene was cloned and sequenced. Colony with phenotype His(+)Mut(+) was isolated by using MD and MM plates and His(+) Mut(+) transformant with high resistance to G418 was screened out by using YPD plate. Using lysate of His(+) Mut(+) colony with high copies of the target gene digested with yeast lyase as the template and 5'AOX1 and 3'AOX1 as the primers, the target fusion gene in chromosome DNA of the constructed P. pastoris engineering strain was detected by PCR. Methanol in BMMY medium was used to induce the target recombinant protein rLipL32/1-rOmpL1/1 expression. rLipL32/1-rOmpL1/1 in the medium supernatant was extracted by using ammonium sulfate precipitation and Ni-NTA affinity chromatography. Output and immunoreactivity of rLipL32/1-rOmpL1/1 were measured by SDS-PAGE and Western blot methods, respectively.Amplification fragments of the obtained fusion gene lipL32/1-ompL1/1 was 1794 bp in size. The homogeneity of nucleotide and putative amino acid sequences of the fusion gene were as high as 99.94 % and 100 %, respectively, compared with the sequences of original lipL32/1 and ompL1/1 genotypes. The constructed eukaryotic expression system was able to secrete rLipL32/1-rOmpL1/1 with an output of 10 % of the total proteins in the supernatant, which located the expected position after SDS-PAGE. The rabbit anti-rLipL32/1 and anti-rOmpL1/1 sera could combine the expressed rLipL32/1-rOmpL1/1.An eukaryotic expression system with high efficiency in P.pastoris of L.interrogans lipL32/1-ompL1/1 fusion gene was successfully constructed in this study. The expressed fusion protein shows specific immunoreactivity, which can be used as a potential antigen for developing a novel vaccine of L.interrogans." @default.
- W2374663397 created "2016-06-24" @default.
- W2374663397 creator A5000893456 @default.
- W2374663397 creator A5011808389 @default.
- W2374663397 creator A5025988922 @default.
- W2374663397 creator A5057374638 @default.
- W2374663397 creator A5062767761 @default.
- W2374663397 creator A5066556669 @default.
- W2374663397 creator A5067128746 @default.
- W2374663397 date "2005-01-01" @default.
- W2374663397 modified "2023-09-23" @default.
- W2374663397 title "[Eukaryotic expression of Leptospira interrogans lipL32/1-ompL1/1 fusion gene encoding genus-specific protein antigens and the immunoreactivity of expression products]." @default.
- W2374663397 doi "https://doi.org/10.3785/j.issn.1008-9292.2005.01.007" @default.
- W2374663397 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/15693121" @default.
- W2374663397 hasPublicationYear "2005" @default.
- W2374663397 type Work @default.
- W2374663397 sameAs 2374663397 @default.
- W2374663397 citedByCount "1" @default.
- W2374663397 countsByYear W23746633972015 @default.
- W2374663397 crossrefType "journal-article" @default.
- W2374663397 hasAuthorship W2374663397A5000893456 @default.
- W2374663397 hasAuthorship W2374663397A5011808389 @default.
- W2374663397 hasAuthorship W2374663397A5025988922 @default.
- W2374663397 hasAuthorship W2374663397A5057374638 @default.
- W2374663397 hasAuthorship W2374663397A5062767761 @default.
- W2374663397 hasAuthorship W2374663397A5066556669 @default.
- W2374663397 hasAuthorship W2374663397A5067128746 @default.
- W2374663397 hasConcept C104317684 @default.
- W2374663397 hasConcept C111829193 @default.
- W2374663397 hasConcept C123894998 @default.
- W2374663397 hasConcept C150194340 @default.
- W2374663397 hasConcept C153911025 @default.
- W2374663397 hasConcept C178790620 @default.
- W2374663397 hasConcept C185592680 @default.
- W2374663397 hasConcept C2777563447 @default.
- W2374663397 hasConcept C40767141 @default.
- W2374663397 hasConcept C55493867 @default.
- W2374663397 hasConcept C86803240 @default.
- W2374663397 hasConceptScore W2374663397C104317684 @default.
- W2374663397 hasConceptScore W2374663397C111829193 @default.
- W2374663397 hasConceptScore W2374663397C123894998 @default.
- W2374663397 hasConceptScore W2374663397C150194340 @default.
- W2374663397 hasConceptScore W2374663397C153911025 @default.
- W2374663397 hasConceptScore W2374663397C178790620 @default.
- W2374663397 hasConceptScore W2374663397C185592680 @default.
- W2374663397 hasConceptScore W2374663397C2777563447 @default.
- W2374663397 hasConceptScore W2374663397C40767141 @default.
- W2374663397 hasConceptScore W2374663397C55493867 @default.
- W2374663397 hasConceptScore W2374663397C86803240 @default.
- W2374663397 hasIssue "1" @default.
- W2374663397 hasLocation W23746633971 @default.
- W2374663397 hasOpenAccess W2374663397 @default.
- W2374663397 hasPrimaryLocation W23746633971 @default.
- W2374663397 hasRelatedWork W2347826495 @default.
- W2374663397 hasRelatedWork W2354537891 @default.
- W2374663397 hasRelatedWork W2355358276 @default.
- W2374663397 hasRelatedWork W2355778255 @default.
- W2374663397 hasRelatedWork W2358194996 @default.
- W2374663397 hasRelatedWork W2366547837 @default.
- W2374663397 hasRelatedWork W2366605413 @default.
- W2374663397 hasRelatedWork W2377593362 @default.
- W2374663397 hasRelatedWork W2383667073 @default.
- W2374663397 hasRelatedWork W2386724379 @default.
- W2374663397 hasRelatedWork W2388429897 @default.
- W2374663397 hasRelatedWork W2391024341 @default.
- W2374663397 hasRelatedWork W2392245231 @default.
- W2374663397 hasRelatedWork W2392719879 @default.
- W2374663397 hasRelatedWork W2406959902 @default.
- W2374663397 hasRelatedWork W2410404115 @default.
- W2374663397 hasRelatedWork W2415029552 @default.
- W2374663397 hasRelatedWork W2415143415 @default.
- W2374663397 hasRelatedWork W3028833262 @default.
- W2374663397 hasRelatedWork W3030263189 @default.
- W2374663397 hasVolume "34" @default.
- W2374663397 isParatext "false" @default.
- W2374663397 isRetracted "false" @default.
- W2374663397 magId "2374663397" @default.
- W2374663397 workType "article" @default.