Words similar to gst-cdc
Example sentences for: gst-cdc
How can you use “gst-cdc” in a sentence? Here are some example sentences to help you improve your vocabulary:
Net1N includes the N-terminal 341 amino acids of Net1 and is sufficient to bind and inhibit Cdc14 [ 11 ] . When GST-Cdc14/His6-T7-Net1N complex anchored to anti-T7 beads was treated with ATP plus increasing amounts of recombinant Cdc5 purified from insect cells, both Cdc14 and Net1N were phosphorylated as indicated by their increased apparent molecular weight.
To assemble Net1N beads with Cdc14 for six reactions (Figure 4A, detection method: Coomassie Blue staining), 105 μl of anti-T7 beads (Novagen) were incubated with 24 μg of His6-T7-Net1N in 100 μl of HEBD for 1 hr, washed four times with HBS + 0.2% n-octylglucoside + 2 mM DTT, and incubated with 6 μg of GST-Cdc14 in 100 μl of HEBD for 1 hr.
GST-T7-Cdc14 (RDB 872) and GST-Cdc14 (RDB 1183) were expressed in E. coli BL21(DE3) pLysS cells, and purified on glutathione-agarose beads (Sigma) as described previously [ 33 ] . His6-T7-Net1(1-341) (Net1N, RDB1153) [ 11 ] was expressed in E. coli BL21(DE3) pLysS cells, and purified on Ni-NTA agarose beads (Qiagen) as described previously for p28 Kix1 [ 34 ] . Purification of His6-Plx1, His6-Plx1(N172), and His6-HA-Cdc5 from baculovirus-infected insect cells followed an earlier protocol [ 24 ] . The concentrations of purified proteins were in the ranges of: Net1N~0.
To test if Cdc5 could dislodge Cdc14 from Net1, we devised a simplified in vitro release assay using GST-Cdc14 and His6-T7-Net1N expressed and purified from bacteria.
p35 was purified as high salt (300 mM NaCl) eluate from the GST-Cdc6 beads.
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