[Characterization and analysis of mitochondrial protein frataxin in Nosema bombycis].
OBJECTIVE
Frataxin protein is a component of Fe-S clusters and closely related to metabolism of mitochondria. We identified an integrity mitochondrial protein frataxin gene (Nbfra), analyzed its phylogenetic relationship, and confirmed the transcriptase activity of Nbfra in N. bombycis.


METHODS
We analyzed the sequence of the second structure, gene location in genome and construction of NJ phylogenetic tree through various bioinformatics software. We constructed recombinant vector pGEX-4T-1-Nbfra, expressed the 36.5kDa recombinant protein in E. coli BL21 (DE3), and then used the protein as antigen to produce its polyantibody in mice.


RESULTS
Nbfra was lack of targeting signal into mitochondria and part of alpha helices in functional domain, and had a synteny character between N. bombycis and E. cuniculi. Phylogenetic trees of Nbfra suggested that the evolutionary position of microsporidia was closely related to that of higher eukaryote, rather than that of other protozoa. The result of western blot suggested the expression and transcription of Nbfra gene in N. bombycis.


CONCLUSIONS
Our results offered the new evidence to analysis the conservation of Nbfra and evolutionary position of N. bombycis, and would support the hypothesis of mitosome in microsporidia.
Hu
Junhua
J
Key Laboratory of Sericulture of Agricultural Ministry, Southwest University, Chongqing 400716, China. dhujh@yahoo.com.cn
Pan
Guoqing
G
Xu
Jingshan
J
Lu
Kun
K
Dang
Xiaoqun
X
Wu
Zhengli
Z
Li
Yanhong
Y
Zhou
Zeyang
Z
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D003001:Cloning, Molecular; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D019143:Evolution, Molecular; D005656:Fungal Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D033862:Iron-Binding Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D051379:Mice; D024101:Mitochondrial Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016818:Nosema / Q000235:genetics; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D000096923:Frataxin
D012321:DNA-Directed RNA Polymerases; D005656:Fungal Proteins; D033862:Iron-Binding Proteins; D024101:Mitochondrial Proteins; D000096923:Frataxin