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Novel insights in the binding of ligands to SMO/StyA, provided by the current protein model, will aid the rational design of mutants with specific, altered enantioselective properties.
The approach discussed by Gonçalves (2007), Goncalves and Resende (2011) says that rather than performing mutation on the entire population, certain mutants with specific mutation rates are introduced in the non-elite section of the population.
Also, the addition of a randomized residue at positions 91 and 93 as well as outer peptide epitopes yielded several mutants with specific activity comparable to the wild-type enzyme with similar outer peptides.
Traditionally, functional genetic studies have been performed by so called forward genetics, in which random gene mutations are generated by induction (e.g., radiation, chemical treatments, and/or insertional mutagenesis), and then mutants with specific phenotypes are identified by breeding and segregation processes (Gao et al., 2008; Lawson and Wolfe, 2011).
Homozygous adult female mutants with specific inactivation of the Nr2f2 in uterine have severely impaired placental formation, leading to miscarriage at days 10 12 of pregnancy [23].
In contrast, lesions in genes encoding transcription factors, receptors, and ligands tend to result in mutants with specific developmental defects in one or a few organ systems [13].
Similar(46)
GST fusion proteins of PI5P4Kγ and PI5P4Kγ+, a mutant with specific activity close to that of the active PI5P4Kα isoform [ 12], were harvested by binding to glutathione sepharose beads (GE Healthcare) and cleaved in situ with 50 units of PreScission protease (GE Healthcare) for 4 h at 4°C.
Furthermore a detailed analysis of the candidate secondary CESAs were performed by functionally characterizing mutants generated with specific artificial microRNA constructs.
In an approach that is akin to phenocopy assays, mutants with altered specific cellular process could be complemented with pathogen effectors and screened phenotypically.
To investigate whether Dkk1 deficiency may have the opposite effect on the specification between SE and NE, we analyzed Dkk1 null mutant embryos with specific markers (Fig. 4L X, Fig. S2D′′ F′′) (Pietila et al., 2011).
Mutant strains with specific deletions of SPI-19 (SG1024 to SG1054) or clpV (SG1034) gene and the concomitant insertion of a Cam-resistance cassette were constructed using the Lambda Red recombination method [23].
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