Cloning and expression of a ferritin subunit for Galleria mellonella.
Ferritin was purified from iron-fed Galleria mellonella hemolymph by ultra centrifugation and FPLC (Superose 6). SDS-PAGE revealed three bands of 26, 30, and 32 kDa. The ferritin 26 kDa subunit cDNA was obtained from RT-PCR using primer designed from N-terminal sequence analysis. 5'-RACE was used to obtain the complete protein coding sequence. The sequence encodes a 211 amino acid polypeptide including a 20 amino acid leader peptide. An IRE (iron-responsive element) sequence with a predicted stem-loop structure was present in the 5'-UTR of ferritin mRNA. Sequence alignment has a sequence identity with Calpodes ethlius (S)(74%), Drosophila melanogaster (50%), and Aedes aegypti (39%). Northern blot analysis indicated that there were 1.5- and 1.75-fold increases in the expression of ferritin mRNA after iron-fed fat body and midgut, respectively. Also, we confirmed that the ferritin mRNA is not expressed in adult ovary and testis. Arch.
Kim
B S
BS
Department of Biology, Korea University, Seoul, Korea.
Yun
C Y
CY
Yeo
S M
SM
Lee
H J
HJ
Kim
H R
HR
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005260:Female; D005293:Ferritins / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D019476:Insect Proteins / Q000235:genetics*; D007501:Iron / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D009036:Moths / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger; D012045:Regulatory Sequences, Nucleic Acid; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution
D018076:DNA, Complementary; D019476:Insect Proteins; D012333:RNA, Messenger; D005293:Ferritins; D007501:Iron