An S. C Technician having acquired S&C Maintainer's Helper seniority will be accorded a S&C Maintainer's seniority date at the time a S&C Maintainer's Helper who is junior to him in seniority, acquires S&C Maintainer's seniority.
An S. C Department employee who first establishes seniority in Agreement 11.8 in the Helper or Apprentice classification, shall also establish a corresponding seniority date in the same classification on the St. Xxxxxxxx Region.
An S. E.I.U.-appointed unit member and the Superintendent or his or her designee shall comprise the Professional Growth Committee.
An S. C Maintenance employee who, in order to protect their seniority, must occupy a bulletined temporary position at a headquarters location over fifty (50) kilometers away from their residence will be allowed a per diem allowance of $37.10 (effective January 1, 2014 $39.10, effective January 1, 2016, $41.10) for each day worked at the location. This per diem allowance will remain in effect for the employee receiving it during the present or subsequent bulletin awards provided:
An S cerevisiae strain (MaV203: pGADGAL) that contains the transcriptional activator protein GAL4 with two amber mutated permissive sites was used for library screening. In the positive selection, because of a GAL4-controlled URA3 gene, cells that can grow in the presence of non-canonical amino acids and in the absence of uracil are the ones carrying active aaRS mutants, which can aminoacylate tRNA with non-canonical amino acids and/or canonical amino acids. Consequently, addition of 5- FOA and removal of the non-canonical amino acids in the negative selection, cells that survived are expressing aaRS specific to the desired analogue because 5-FOA is converted to a toxic product with the expression of URA3 gene. In this way, more than 20 unnatural amino acids have been added to the yeast genetic code in response to UAG codon. (Liu and Xxxxxxx, 2010) The addition of unnatural amino acids to the genetic code provides another possibility in the evolution of proteins with novel properties. However, in the approach of expanding genetic code in yeast, the scope of current orthogonal pairs is still limited to mutant E. coli tRNATyr/TyrRS, a human initiator tRNAfMet/E. coli GlnRS and M. barkeri tRNAPyl/PylRS pairs. (Chin et al., 2003; Xxxxxxx et al., 2010; Xxxxx et al., 2001). In addition, non-canonical amino acids have been inserted into proteins only in response to the amber (UAG) codon. Therefore, in Chapter 2 of this thesis, we describe the methodology of constructing an orthogonal Mycoplasma genetalium tryptophanyl-tRNA synthetase (MgenTrpRS)/MgentRNATrp) pair for incorporation of non-canonical amino acids in S. cerevisiae cell in response to opal (UGA) codon. In chapter 3, a library selection scheme for the identification of mutant MgenTrpRS variants with altered selectivity for the non-canonical amino acids, 5-hydroxy-tryptophan and 5-methoxy- tryptophan, is investigated and the spectroscopic properties of enhanced cyan fluorescent protein (ECFP) containing these non-canonical amino acids in the chromophore is described.
