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To determine if ERK/MAPK requires the D-domain for efficient cyclin D1 phosphorylation at Thr286, we performed in vitro kinase assays using a complete deletion of the D-domain (ΔD) from the GST-C-terminal cyclin D1 fusion protein.
Trees were rooted at midpoint, using a complete deletion or pairwise deletion, the Poisson correction option, and 500 bootstrap replicates.
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Whereas we used a complete deletion (null) allele of uvrD, they used an allele that encodes a truncated 230 amino acid UvrD protein, which may still contain ATP binding and other activity.
For our test of this method we used mice with a complete deletion of the BChE catalytic domain [9].
BY384 (MATa leu2Δ1 lys2Δ202 trp1Δ63 ura3-52 his3Δ200) was used to generate a complete deletion of the tRNASerGCU gene family.
The NJ tree was constructed using MEGA 4.1, and Maximum Composite Likelihood distance [ 47] was used and gaps were treated as a complete deletion.
MP analysis was performed with a heuristic search using close neighbor interchange (level = 1), a branch-swapping method with initial trees generated by random addition (10 replications); a complete deletion option was used to treat gaps/missing data.
A complete deletion option was used to calculate the distances and the bootstrap method with 1000 replicates was employed for estimating the standard errors.
We successfully used these lines along with one that carries the FLP recombinase to generate a complete deletion allele of the gurken gene, called grkΔFRT.
In the second round of allelic exchange, for example to create a complete deletion, the temperature sensitive plasmid containing only the homologues flanking regions is used (Fig. 2).
The million-SNP array indicated a complete deletion of about a third of both copies of a gene known to be involved in the digestion of certain starches.
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