Expression cloning and characterization of a pupal cuticle protein cDNA of Galleria mellonella L.
Epidermal mRNA of freshly ecdysed pupae of Galleria mellonella was used to establish a cDNA library in phage lambda gt11. A cDNA clone was isolated by means of a pupal cuticle protein (PCP) specific antibody. Nucleic acid sequence data show an insert of 1212 bp with an open reading frame of 1062 bp. The presence of start, stop, and polyadenylation sites suggests, that this insert represents a full length transcript. Conceptual translation resulted in a protein of 353 amino acids including a signal peptide. The final processed protein product is rich in alanine and charged amino acids like glutamic acid. It has a calculated pI of 4.19 and a molecular mass of 34.272 kDa. In hybrid selection/in vitro translation and in vitro transcription/translation experiments a translational product of 54 kDa was synthesized. The difference between apparent and calculated molecular mass is thought to be due to the relatively high amount of glutamic acid residues clustered in two regions. The developmental profile of expression of the corresponding gene was analyzed by northern blot hybridization using a cDNA probe. The time course of expression is coincident with developmentally regulated metamorphic changes in the integument.
Kollberg
U
U
Theodor-Boveri-Institut für Biowissenschaften, Lehrstuhl Zoologie I, (Zell- und Entwicklungsbiologie), Würzburg, Germany.
Obermaier
B
B
Hirsch
H
H
Kelber
G
G
Wolbert
P
P
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D018507:Gene Expression Regulation, Developmental; D008969:Molecular Sequence Data; D009036:Moths / Q000235:genetics*; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics*; D011679:Pupa / Q000235:genetics; D012333:RNA, Messenger; D014158:Transcription, Genetic
D018076:DNA, Complementary; D011506:Proteins; D012333:RNA, Messenger