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Peptide activates USP45. 39 / 39
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"On the one hand, it may be because the fusion of the initial signal peptide with Usp45 signal peptide would affect the folding of the signal peptide, which in turn affects its function."
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"Whole cell ELISA assay showed that pSD1nuc which had the combination of P promoter with USP45 signal peptide gave very low signals in the ELISA assay, although fluorescent signals were still detectable in immunofluorescence assay (Fig. 7)."
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"A cassette consisting of USP45 fused with LEISSTCDA propeptide to increase secretion efficiency, pNZ8048 multiple cloning sites (MCS) modified to include more RE sites, pNZ8048 terminator (T), and TP901–1 bacteriophage attachment site (attP) was synthesized by Integrated DNA Technologies (Singapore) and denoted as attP cassette.The vectors constructions strategy was divided into four phases; Phase I) Construction of PZatt cassette composed of the P promoter, USP45 signal peptide, LEISSTCDA propeptide and attP sequence; Phase II) Construction of the secretion integrative vectors (pS1–4); (a) Plasmid circularisation by incorporation of erythromycin resistant gene and E. coli replication origin into the PZatt cassette, denoted as pS1; (b) Replacement of P promoter with P promoter to produce pS3 plasmid; (c) Replacement of USP45-LEISSTCDA fusion with SPK1 signal peptide into each pS1 and pS3 plasmids to produce pS2 and pS4 plasmids (Additional file 3: Figure S1); Phase III) Construction of the surface display integrative vectors (pSD1–4) by incorporation of PrtP anchor domain into each secretion integrative vector; Phase IV) Cloning of reporter gene, nuclease (Additional file 4: Figure S2)."
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"The present investigation included cloning the BAB1-0278 gene fragment, which contains 237 base pairs coupled with the Usp45 signal peptide, into the pNZ8148 vector using K pnl and X bal restriction e[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"Substantially higher levels of the fusion proteins (four- to six-fold) were secreted by the clones possessing Lactobacillus brevis SlpA signal peptide than by those possessing L. lactis Usp45 signal peptide."
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"On the other hand, the pS2 plasmid was constructed by replacing USP45 signal peptide with SPK1 using EcoRV and KpnI RE."
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"To obtain BaMTGase into the medium, the DNA encoding Usp45 signal peptide (SPusp45) was fused to the B. amyloliquefaciens mtg gene to direct the secretion."
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"Secretion using Usp45 signal peptide generally gave satisfactory efficiency."
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"For example, using Usp45 signal peptide, a Bacillus nattokinase was almost completely secreted to the medium ( Liang et al., 2006 )."
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"Compared to the control which utilizes Usp45 signal peptide, the new signal peptide allowed much higher secretion of mIL-12, 185 pg/ml versus 40 pg/ml ( Fernandez et al., 2009 )."
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"The 3243 bp of the 4,3-α-GTase enzyme was successfully cloned into the pLEB124 vector by adding the usp45 signal peptide to its N-terminal region."
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"The usp45 gene encodes the major extracellular protein Usp45 from L. lactis MG1363 and chimeras of the 27-amino-acid Usp45 signal peptide (SP usp45 ), fused to mature proteins, have been used to drive[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"The subsequent amino acid sequence-mass spectrometry sequencing of protein content in the upper and bottom TRAIL bands, revealed that in both bands TRAIL protein occurs beyond any doubt (Table 1) and in both TRAIL fractions the usp45 leader peptide (27 aa) was cut off."
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"To calculate the efficacy of hsTRAIL secretion, when pNZ8124 plasmid vector with sequence for usp45 leader peptide was used, we performed ELISA and quantified the concentration of the protein both in crude supernatants and cell lysates after 4 h upon the nisin-induction."
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"E. faecium T136 was used as the source of entA and entiA , whereas L. lactis MG1363, E. faecium P13, and E. hirae DCH5 were used as the source of the Usp45 signal peptide (SP usp45 ), the EntP signal [MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"However, fusion with Usp45 signal peptide (SP Usp45 -LysL) weakened LysL activity, likely due to incomplete signal peptide cleavage during secretion."
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"Alternatively, Le Loir et al. (2001) described a strategy using a combination of the Usp45 signal peptide and a synthetic propeptide to improve the secretory production of proteins."
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"The 87-bp Usp45 signal peptide was amplified from the genomic of L. lactis MG1363 using primers 5 and 6 and was cloned upstream of the CGTase gene in pNZ:G1-CGT to obtain plasmid pNZ:Usp45-CGT ( Table[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"The fusion of the Usp45 signal peptide with the CGTase gene lacking the native signal peptide (G1) yielded the vector pNZ:Usp45-CGT."
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"The G1 and M5 signal peptides are 29-amino acids long, whereas the L. lactis Usp45 signal peptide is 27-amino acids long."
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"The Usp45 signal peptide contains a basic N-terminus (+3) while the G1 and M5 signal peptides have acidic N-termini (−1) ( Table 4 )."
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"The Usp45 signal peptide has been engineered via random mutagenesis in its nucleotide sequence as well as in its peptide sequence for enhanced secretion ability as well as enhanced translation efficie[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"An amino acid sequence analysis revealed that the G1 and M5 signal peptides are 29 amino acid long, whereas the L. lactis Usp45 signal peptide consists of 27 amino acids."
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"The USP45 signal peptide has been employed to enable the secretion of target proteins by L. lactis [32]."
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"The Usp45 signal peptide is added to the N-terminus of the target protein to facilitate its secretion by L. lactis [28]."
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"In the study by [26], the pNZ8124:sip vector containing the lactococcal Usp45 signal peptide sequence (SP usp45) fused to the PnisA promoter was successfully constructed, allowing the target protein to be expressed extracellularly by L. lactis."
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"[17] reported the successful utilization of the Usp45 signal peptide for the extracellular expression of the Helicobacter pylori Lpp20 antigen using L. lactis."
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"Other researchers, such as [15, 22, 25, 30-32, 34, 35, 37] have also proven that the usp45 signal peptide can be used to express extracellular recombinant proteins in L. lactis."
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"The food-grade expression vector pNZ-SECF1S1 harboring the nisin-inducible promoter (PnisA) and Usp45 signal peptide was constructed."
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"For anchoring EG95 protein to the cell surface, we utilized Usp45 signal peptide at the N-terminal and the M6 cell-wall anchor at the C-terminal of the protein, and our results showed that EG95 could effectively be exhibited at the cell wall of L. lactis (Fig. 4)."
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"Our data confirmed that the use of sp6 anchor in addition to usp45 signal peptide and interval sequence can efficiently display LTB on the surface of L. lactis."
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"The capacity of this system to produce cytoplasmic and secreted proteins was initially tested using the coding sequence of staphylococcal nuclease ( nuc ) fused or not to the lactococcal Usp45 signal [MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"To generate a L. reuteri strain capable of secreting ShK-235, we codon-optimized the gene for ShK-235 for expression in L. reuteri, fused the modified usp45 signal peptide (21, 22) sequence to the 5′ end, and cloned it into the vector pSIP411 (23) resulting in plasmid pLL01."
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"The psaA gene was cloned into the pT1NX vector PstI site, which produced a fusion of a truncated usp45 signal peptide carried by the vector to the PsaA sequence (Fig. 1), under the control of a constitutive promoter."
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"The quantity of secreted barnase from L. lactis (pNZ8048- A45Ban Δ sp -Bas ) was 5.6 times higher than from L. lactis (pNZ8048- A45Ban-Bas ) implying that a Usp45 signal peptide enabled a significantl[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"P1 represents the constitutive promoter; SD, Shine-Dalgarno sequence; SS, first codons of the Usp45 signal peptide fused to the psaA gene.Fig."
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"As for the surface display integrative vector, all systems developed were comparable with the exception of the combination of P 170 promoter with USP45 signal peptide which gave very low signals in whole cell ELISA."
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"The 3243 bp of the 4,3 α-GTase was successfully cloned ( Fig. 1 b) into the pLEB124 vector by adding the usp45 signal peptide to its N-terminal part."
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"Extracellular targeting using only the Usp45 signal peptide sequence is more advantageous."
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