Molecular cloning and characterization of the B subunit of a vacuolar H +-ATPase from the midgut and malpighian tubules of Helicoverpa virescens
Using the polymerase chain reaction (PCR) a 0.8-kb product was amplified from cDNA made from the midgut and Malpighian tubules of fifth instar larvae of Helicoverpa virescens. This 0.8-kb PCR product was then used to isolate a clone of the B subunit of the V-type ATPase from a cDNA library made from...
Gespeichert in:
Veröffentlicht in: | Archives of biochemistry and biophysics 1991-11, Vol.291 (1), p.92-99 |
---|---|
Hauptverfasser: | , |
Format: | Artikel |
Sprache: | eng |
Schlagworte: | |
Online-Zugang: | Volltext |
Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
Zusammenfassung: | Using the polymerase chain reaction (PCR) a 0.8-kb product was amplified from cDNA made from the midgut and Malpighian tubules of fifth instar larvae of
Helicoverpa virescens. This 0.8-kb PCR product was then used to isolate a clone of the B subunit of the V-type ATPase from a cDNA library made from the same tissues. The cDNA clone encodes for a protein of 55 kDa which shows very high amino acid homology to other known B subunits of V-type ATPases. The transcript size of the B subunit in the midgut of
H. virescens was 2.3 kb, and a transcript of identical size was also detected in the Malpighian tubules. Northern blot analysis revealed the presence of a homologous transcript of 2.6 kb in the midgut of
Manduca sexta and PCR analysis also confirmed the presence of such a transcript in the Malpighian tubules and the nervous system of
M. sexta, and in the midgut Malpighian tubules of
Culex quinquefasciatus. The presence of the V-type ATPase in the Malpighian tubules of lepidopteran insects suggests that the transport of ions across the cell membrane in this tissue is also probably driven by a similar process as that observed in the midgut of these insects. |
---|---|
ISSN: | 0003-9861 1096-0384 |
DOI: | 10.1016/0003-9861(91)90109-V |