title: cDNA cloning and sequence analysis of Musca domestica antifungal peptide-1 (MAF-1).
abstract: OBJECTIVE
  To clone the cDNA sequence of Musca domestica antifungal peptide-1 (MAF-1) and analyze the amino acid sequence of MAF-1 by bioinformatics method.
  
  
  METHODS
  Based on the primer designed according to the N-terminal amino acid sequence of MAF-1, the cDNA and amino sequence of MAF-1 were obtained by the methods of RACE and NestPCR. The accuracy of the experiment was confirmed by RT-PCR The characteristic of the sequence was analyzed by bioinformatics software.
  
  
  RESULTS
  The length of the cDNA sequence of MAF-1 was 568 bp by 3'RACE, including an open reading frame (ORF) of 441 bp length and 3'UTR of 127 bp. It was a novel sequence with the submission number of HM178948 in GenBank since none homology was found when compared with other sequences by Blast. Added with the 9 amino acids that were not used to design primer, the whole sequence of MAF-1 was 156 amino acids conferred from its cDNA. 139 bp cDNA sequence was obtained by 5'RACE and the result was consistent to 3'RACE. The result of RT-PCR showed the cDNA of MAF-1 mature peptide was accurate. The bioinformatics analysis deduced that the theoretic molecular weight and isoelectric point of the whole protein sequence of MAF-1 gene were similar to those detected. The ExPASy illustrated that the MAF-1 gene had a signal peptide. There were abundant a-helix in it, the domain located between the 128 and 153 amino acid residuals. Subcellular analysis showed MAF-1 was almost in the nucleus. Predict Protein found two protein kinase C phosphorylation sites and one N-myristoylation site, and predicted that it was not a globular protein. In the end, the three dimension image of MAF-1 was set up with 3D-pssm of ExPASy.
  
  
  CONCLUSIONS
  The cDNA sequence and the amino acid sequence of MAF-1 have been obtained and analyzed successfully.
authors.affiliation: Department of Parasitology, Guiyang Medical College, Guiyang 550004, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Fu
authors.forename: Jianwei
authors.initials: J
authors.lastname: Wu
authors.forename: Guo
authors.initials: G
authors.lastname: Guo
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D019295:Computational Biology; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D006793:Houseflies / Q000235:genetics*; D019476:Insect Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics*; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D019476:Insect Proteins; D010455:Peptides