Cloning and expression of a ferritin subunit forGalleria mellonella

Autor: Sung Moon Yeo, Beom Su Kim, Hye Jeong Lee, Hak R. Kim, Chi Young Yun
Rok vydání: 2001
Předmět:
Zdroj: Archives of Insect Biochemistry and Physiology. 47:8-17
ISSN: 1520-6327
0739-4462
DOI: 10.1002/arch.1030
Popis: 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. Insect Biochem. Physiol. 47:8–17, 2001. © 2001 Wiley-Liss, Inc.
Databáze: OpenAIRE