Cloning of cDNA for vitellogenin of Athalia rosae (Hymenoptera) and characterization of the vitellogenin gene expression.
Athalia rosae (Hymenoptera) was previously shown to have two vitellins (L-Vn and S-Vn) and the two corresponding vitellogenins (L-Vg and S-Vg). A cDNA expression library was constructed from poly(A)+ RNA prepared from adult female fat body cells, and was screened for the vitellogenins by using antisera against the L- and S-Vn, respectively. Examinations of cloned cDNAs show that the vitellogenin gene is transcribed as a single unit, with the 5'-terminal site coding for the S-Vg and the 3'-terminal site for the L-Vg. Nucleotide sequence at the 5'-end suggests the presence of a 16 amino acid-long signal peptide. Deduced amino acid sequence following the signal peptide shows a complete match with up to the 28 N-terminal amino acid sequence determined on S-Vn. The S-L Vg boundary with deduced amino acid sequence matching with 6 N-terminal amino acid sequence determined on L-Vn is also detected. Northern blot hybridization analysis shows that the vitellogenin gene is expressed in the female fat body as a single 6.5 kb mRNA but not in the ovary, and not in the male fat body. Western blot analysis detects a large precursor polypeptide, reacting with the anti-L-Vn and S-Vn antisera, in the adult female fat body.
Kageyama
Y
Y
Department of Biology, Faculty of Science, Kobe University, Japan.
Kinoshita
T
T
Umesono
Y
Y
Hatakeyama
M
M
Oishi
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005216:Fat Body / Q000737:chemistry; D005260:Female; D015870:Gene Expression; D015723:Gene Library; D017344:Genes, Insect / Q000235:genetics*; D006927:Hymenoptera / Q000235:genetics*; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011498:Protein Precursors / Q000032:analysis; D021382:Protein Sorting Signals / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D017421:Sequence Analysis; D017422:Sequence Analysis, DNA; D014819:Vitellogenins / Q000737:chemistry / Q000235:genetics*
D018076:DNA, Complementary; D011498:Protein Precursors; D021382:Protein Sorting Signals; D012333:RNA, Messenger; D014819:Vitellogenins