In the period leading to June 30, we finished obtained raw data from transcription assays reporting function of CLas transcription factors. We analyzed results from RNA transcription assays on the Sm (deletion) visRvisN mutant carrying a plasmid with CLas visNR. The overexpressed CLas visNR caused upregulation of 38 genes with a cut-off of 1.5X. Among these, most dramatically changed was expression of rem (SMc03046, a response regulator for motility during exponential phase growth) and a number of flagellar synthesis genes which went up by 4 to 8X. We also isolated, assayed and analyzed genes responsive to the CLas lsrB gene. The LsrB proteins are LysR-type transcriptional regulators. The S. meliloti ortholog of LsrB protein has previously been shown (Tang and Cheng, 2013) to function in symbiosis and to regulate the lrp3-lpsCDE operon. Our deletion mutant of Sm lsrB grows poorly and is highly susceptible to destruction by the detergent DOC. The cloned CLas lsrB gene partly complements this function, to make Sm lsrB mutants a bit more resistant to DOC. However, our transcription results indicated that CLas LsrB does not function well in S. meliloti as a transcription factor. Specifically, expression of lrp3-lpsCDE was unchanged by the presence of CLas LsrB. On a global level, CLas LsrB only changed expression of 4 genes, and by a maximum of 1.2 fold either up or down Transcript analysis for the CLas CtrA regulator expressed in wild type S. meliloti showed that CLas CtrA upregulates a number of cell-division related genes including minCDE (2 to 3X change). Several genes related to native CtrA regulation were down-regulated by CLas CtrA protein. This result was unexpected, given that no genes showed decreased expression due to the native Sm ctrA gene. The phrR regulatory gene exists in two copies in S. meliloti, with the two proteins (PhrR1 and PhrR2) being 56% identical to each other. PhrR1 protein is 59% and PhrR2 is 48% identical to the CLas PhrR protein. As background for the CLas expression studies, a S. meliloti double mutant of phrR1 and phrR2 genes had to be constructed. This mutant (or even the phrR1 single mutant) grew very poorly. The cloned CLas phrR gene did not restore growth to the S. meliloti mutant, and when expression of CLas PhrR protein was induced with IPTG, it somewhat inhibited growth of the wild type control as well; for example, the cells expressing CLas PhrR protein yielded smaller colonies. The overexpressed PhrR transcription factor did not induce high levels of any gene (maximum upregulation was 1.2X) although it had slightly stronger negative effects on a small number of gene targets. Assembling all the data, we began to select which target genes would be used to construct chromogenic (lacZ) and fluorescent (GFP) fusions for the robotic assay.