Sequence-replacement mutants were generated either by ligation of a double-stranded oligonucleotide sequence into a linearized construct lacking the sequence to be replaced or by performing PCR with primers that contained sequence corresponding to the replacement sequence at their 5′-end (see Supplementary Figure S1 at http://www.biochemj.org/bj/456/bj4560139add.htm).htm
For the analyses of inhibitor sensitivity, we focused on a structurally well-defined kinase domain (KD) of FGFR1 (Mohammadi et al., 1996, Bae et al., 2010, Norman et al., 2012), with the gatekeeper V561M variant and Y563C variant (corresponding to the replacement found in FGFR4) generated in the context of FGFR1.
The effect on durability corresponding to the increasing replacement of fly ash was evaluated, and the optimum value of fly ash replacement was recommended.
The recombinant R65 [45] differs from the native isolate [44] by 4 nucleotide exchanges in the PB2 gene (DQ464357): correspondingponding to the amino acid replacement L329F, in the PB1 gene (DQ464361): and03G1604TG1604T corresponding to the amino acid replacement S527V, and in the HA gene (DQ464354): C1737T in non-coding region.
For the range of 0 45% for the level of fly ash replacement, the rebound value, the ultrasonic velocity, the elastic modulus and the compressive strength all reached their maximum value corresponding to the fly ash replacement level of 15%.
The hyg specific probe 1026-bpp) resulted in no hybridizing bands with SalI-digested genomic DNA from wild type parasites and Ld GLOI::NEO (+/n) clones, while a single band of ∼6.8 kb, corresponding to the hyg based replacement fragment was detected in L. donovani heterozygous clones, Ld GLOI :: HYG (+/h) (Fig. 3A).
The metapopulation was assumed "genetically extinct" when the average offspring survival W(t ) dropped below the threshold W* = F-1, where F was the average basic fecundity (corresponding to the theoretical population's replacement rate in the absence of genetic load).
For each quadrant, using probability laws, we predicted the number of shared singletons when a virtual community is created by randomly drawing a number equal to the encountered number of singletons in each quadrant 4 times (corresponding to the 4 quadrants studied) with replacement from a pool of the expected number of singletons in that quadrant.
The analysis of these curves shows that the peak strain corresponding to the maximum stress increases when increasing the replacement ratio.
Since the segment tree maintains the longest interval at the root node, whenever the buffer overflows, we simply pick up the data pages corresponding to the longest interval as the optimal replacement plan.
To test how the motor domain of MCAK interacts with the C terminus at the molecular level, we co-crystallized and determined the structure of the motor domain bound to a chemically synthesized peptide corresponding to the CT domain (709QLEEQASRQISS720) using molecular replacement to a resolution of 3 Å with good stereochemical parameters (Table 1, Figure 3A).
