Transcript analysis of the genes encoding aminopeptidase N and alanine aminotransferase, two enzymes involved in protein turnover, in the Pacific oyster, Crassostrea gigas.
Molecular probes have been developed to detect aminopeptidase N (ApN) and alanine aminotransferase (ALAT) transcripts in the Pacific oyster Crassostrea gigas. Degenerate primers were designed using ApN and ALAT sequences stored in the EMBL database. Amplification of C. gigas genomic DNA using these primers resulted in amplification of a 344-bp ApN fragment and a 530-bp alanine aminotransferase fragment. The deduced amino acid sequence of the ApN fragment displayed 75 and 73% identities with sequences of ApN from human and mouse, respectively. The deduced amino acid sequence of the ALAT fragment displayed 57% identity both with human and rat ALAT. An ApN transcript of approximately 3.1 kb was detected by northern blotting in larvae and in adult digestive gland and gonadal tissue. No transcript was detected in adult adductor muscle. An ALAT transcript of approximately 2 kb was similarly detected in larvae and in adult gonadal tissue, but was undetectable in adult digestive gland and adductor muscle. Transcript detection employing RT-PCR demonstrated low-level expression of both ApN and ALAT in all studied tissues, in both larvae and adults.
Donald
K M
KM
Plymouth Marine Laboratory, Prospect Place, Plymouth, Devon, PL1 3DH, UK. kdon@wpo.nerc.ac.uk
Hawkins
A J
AJ
Smerdon
G R
GR
D000410:Alanine Transaminase / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D018826:CD13 Antigens / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D017931:DNA Primers / Q000737:chemistry; D018507:Gene Expression Regulation, Developmental; D008969:Molecular Sequence Data; D010125:Ostreidae / Q000201:enzymology*; D012333:RNA, Messenger / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution
D017931:DNA Primers; D012333:RNA, Messenger; D000410:Alanine Transaminase; D018826:CD13 Antigens