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Example sentences for: linkers
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This is analogous to the use of the bovine pancreatic trypsin inhibitor for internal insertions [ 21 ] . The transposons described here could potentially be improved upon by optimizing the length and flexibility of the linkers between the target and the GFP.
RNA was reverse transcribed into cDNA using random hexamer primers in the presence of 5-aminoallyl-dUTP; Cy3 and Cy5 dyes were covalently coupled to the incorporated aminoallyl linkers in a subsequent labeling reaction.
Two synthetic transposons have been reported that can produce GFP fusion proteins [ 9 10 ] . The design of these transposons included additional protein domains or linkers between the GFP and the target protein, however, and little is known about how many of the resulting proteins continued to function.
Sodium salts of each compound were titrated into assays of purified Fhit [ 16 ] with 1.8 μM fluorescent substrate, Appp-S-(4-4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacine-3-yl)methylaminoacetyl (ApppBODIPY) [ 10 ] , and competitive K i values were obtained by calculating the inhibitor concentration-dependence in reduction of k cat / K m (apparent) for substrate hydrolysis [ 10 ] . As shown in Table 1, inhibitors 2b and 3b with two-carbon diol linkers were either only as inhibitory as the natural substrate 1 or substantially less so.
We reasoned that a Tn 5 transposon [ 11 12 ] could be designed that would generate GFP fusion proteins with relatively short linkers (~7 amino acids) between the GFP and the host protein analogous to GFP fusion proteins that have already been shown to function [ 6 8 ] . To test this approach, we targeted the G protein subunit α s and the glutamate receptor subunit GluR1.
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