Puromycin Gfp Fusion

le the stat1 pseudogene lay immediately downstream of stat1b, suggestive of a local duplication event. Only a MedChemExpress 193022-04-7 single gene showed conserved synteny with stat6 while no synteny conservation was evident PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/22189597 for stat2. However, the identity of the latter was confirmed by the presence of a KYLK motif in the encoded protein that is unique to STAT2, as well as a similar splice structure. Conserved synteny was also evident between various STAT gene clusters, such as the GLS-, MYO1- and NAB-related genes between the STAT1/STAT4 cluster and STAT2/STAT6 cluster as well as HSD17B-related genes between the STAT3/STAT5 cluster and STAT2/STAT6 cluster. Phylogenetic analysis suggested two distinct STAT sub-families, one containing vertebrate STAT1, STAT2, STAT3 and STAT4 along with sea squirt stata, and one containing vertebrate STAT5 and STAT6 along with sea squirt statb. This distinction was also supported by the alternative splice structure in the region encoding the coiled-coil and DNA binding domains between these two sub-families. SHP evolution Single SHP homologues have been identified in fruit fly and in sea squirt , whilst mammals possess two family members . Analysis of vertebrate genomes revealed three SHP homologues in zebrafish, with expression confirmed by RT-PCR and the presence of corresponding EST sequences in each case, as well as related genes in several tetrapods, including chicken and toad. Phylogenetic analysis identified two of these as orthologs of mammalian SHP1 and SHP2, confirmed by conserved synteny relationships for shp1 and shp2 , and conservation of splicing structure, functional domains and motifs, including C-terminal tyrosines. Phylogenetic analysis identified a distinct third clade related to both SHP1 and SHP2, including a conserved splice site structure that was named SHP3. Analysis of mammalian genomes identified a SHP3 pseudogene with flanking genes showing conserved synteny to zebrafish shp3, suggesting selective loss of this SHP family member along the mammalian lineage. PIAS evolution There is a single PIAS gene in both fruit fly and sea squirt , whilst there are four PIAS members in mammals . Bioinformatic analysis revealed the presence of four pias genes in zebrafish, the expression of which were confirmed using RT-PCR. Phylogenetic analysis indicated that these represented piasx and piasy orthologs and two pias1 paralogs, pias1.a and pias1.b, with no pias3 ortholog present. The identities of pias1.a, pias1.b and piasy were confirmed by conserved synteny to their mammalian counterparts, including multiple common genes across both pias1 genes with additional genes or gene for pias1.a and pias1.b, respectively. There were also two genes showing conserved synteny to piasy, although there were none for piasx between zebrafish and humans. Japanese pufferfish had the same pias complement as zebrafish, whilst African clawed frog possessed the same pias complement as mammals, suggesting a teleost-specific Evolution of JAK-STAT Pathway Components absence of pias3. Each of the PIAS functional domains, including the SAP, PINIT, RLD, AD, and S/T-rich were conserved in teleost PIAS1 and PIASx homologues. Like their mammalian counterparts, teleost PIASy proteins lack both the AD and the S/T-rich region. Despite extensive searches, additional exons containing the leader sequence for zebrafish pias1.b and piasx were not found. SOCS evolution In contrast to other JAK-STAT pathway components, which exist as single members in inver