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Expression of TetR-mCerulean and LacI-mCherry (Rizzo et al., 2004; Shaner et al., 2004) was by inserting fusion genes, regulated by the lac promoter, into galK or leuB (Supplemental Experimental Procedures).
Because hemifusion requires that fusion peptides of 3 4 adjacent trimers insert into the target membrane, the apparent rate constant for hemifusion relates to a different probability that of inserting fusion peptides from several neighboring HAs.
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Similarly, the surface display vector for the RML gene (pPIC9K-KNS-Rml) was obtained by inserting the fusion gene RML-α-agglutinin into the plasmid pPIC9K.
(GS 8 insert fusion protein variants were expressed in E. coli NiCo21 DE3 RIPL cells.
When gB is activated, it is thought to insert its fusion loops into the target cell membrane and undergo a dramatic conformational change, pulling viral and cellular membranes together and driving fusion [6].
The "classical" postendocytosis fusion pathway depends on acidic pH to induce a conformational change in the viral envelope (E) protein to expose and insert the fusion loop into the endosomal membrane [ 17– 17].
Once, it was all about getting a logo in at the start or end of a game, but the latest technologies from specialist agencies like Massive and Double Fusion involve inserting billboards, videos and even 3D objects into virtual environments.
Co-author Mehboob Hussain explains that they tested for fusion by inserting male marrow cells into females whose beta cells contained an inactivated form of GFP.
Finally, the researchers report in the 18 January issue of Science, they recreated Isabella's preleukemic stem cells by inserting a TEL-AML1 fusion gene into cord blood stem cells with the same surface markers as those that were seen in Isabella.
pBF24C was constructed by amplification of the ermE gene from S. erythraea BIOT-4480 using the primers PBF24D-for and PBF24D-rev and inserting it by In-Fusion cloning into pBF24B cut with XbaI.
All the target genes were successfully inserted into the fusion protein vectors and expressed at high level from at least one of the fusion vectors.
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