Cloning Of Bacillus Subtlis YlyA’ And Bioinformatics Analysis

Study of transcription factor is a key area to investigate the regulation of gene expression. COG database and Smith-Waterman report for YlyA predicted that the hypothetical protein YlyA function as a DnaK suppressor. The protein database Pfam and UniProtKB/Swiss-Prot predicted that YlyA contains a C4type zinc finger structure which resembles the transcription factor DksA from E. coli. All these structural and functional information indicated YlyA is a possible transcription factor which can provide new target for drug design. What’s more, sequence blast result between YlyA and known proteins in COG suggested that YlyA produced significant alignment with52proteins from30sets of genomes, this suggested that YlyA plays the same function as these proteins. The majority of microbes producing these proteins are pathogens causing severe infectious disease including E.coli O157.H7, Salmonella typhimurium, Yersinia pestis, Vibrio cholerae, bacillus anthrax, Pasteurella multocida, Brucella melitensis et al. Research of YlyA, the putative transcriptional factor, is expected to provide new antibiotic target for the treatment of above diseases.I ylyA fragment was amplified from Bacillus subtilis chromosome DNA of standard strain Bs168by PCR using primer pairs designed according to the ylyA sequence published in the subtlist database, the sequencing result showed that the amplified ylyA is different from that of subtilist in six nucleotides located in5’terminal of the sequence. Then the new ORF was found downstream of the original sequence, the new one was named as ylyA’which use the same stop codon as original ylyA. The full-length ylyA’is375bp in length and encodes124amino acids, there are only4amino acids located in N terminal that differs from YlyA.II The bioinformatics analysis for YlyA’ showed that:YlyA’ is basically a globular protein with a molecular weight of14.1KDa, which is negatively charged in the cell’s physiological conditions. The protein doesn’t contain transmembrane helices but hydrophobic regions. A DksA domain and a zf-dskA-traR type zinc finger were found exist in the protein. Therefore, theoretically YlyA’ can bind to the DNA by via of its zinc finger structure, and has a resemble function as transcription factor DksA of E.coli which is a DnaK suppressor in the signal transduction process. Protein-protein blast result confirmed the above predictions, so the computer analysis strongly suggested that YlyA’ is a possible transcription factor. The present analysis will provide a theoretical foundation for future further studies on overproduction, subcellular localization and interaction with RNAP.