ISSN:
1432-136X
Keywords:
Key words Amylase regulation
;
Digestive enzymes
;
Oyster
;
Feeding stimulus
;
Abbreviations bp base pair
;
cDNA copy DNA
;
CCK cholecystokinine
;
EDTA ethylenediaminetetraacetic acid
;
IGF insulin growth factor
;
MOPS 3-[N-morpholino] propane sulfonic acid buffer
;
PCR polymerase chain reaction
;
pfu plaque forming unit
;
SDS sodium dodecylsulfate
;
SSC sodium citrate buffer
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
,
Medicine
Notes:
Abstract A Crassostrea gigas digestive gland copy DNA (cDNA) library constructed in the λ phage ZapII (Stratagene, La Jola, USA) was screened with an amylase heterologous probe. To get access to the complete cDNA, a polymerase chain reaction extension was conducted using DNA extracted from the phages. The complete cDNA sequence is 1688 base pairs (EMBL = Y08370). The deduced protein sequence is 519 aminoacids long with a 19 aminoacid signal peptide. Similarity with Pectenmaximus amylase is 72%. A 3-day nutrition experiment with a cyclic algal food supply was carried out. Amylase enzyme activities and mRNAs were individually measured on five animals, nine times a day. Messenger RNAs were quantified by dot hybridization using the previously characterized cDNA as probe. Variation of amylase mRNA was observed, in relation with the level of activity of the enzyme. Coordinated changes in RNA and enzyme levels suggested a possible transcriptional regulation of amylase in C. gigas as in vertebrates.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/s003600050003
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