Cloning and sequencing hybrid striped bass (Morone saxatilis x M. chrysops) transforming growth factor-β (TGF-β), and development of a reverse transcription quantitative competitive polymerase chain reaction (RT-qcPCR) assay to measure TGF-β mRNA of teleost fish

A transforming growth factor (TGF)-β was isolated and cloned from hybrid striped bass (Morone saxatilis x M. chrysops) anterior kidney mononuclear cells. This isolate (Genbank accession number AF140363) contains an open reading frame of 1146 bases coding for a 382 amino acid protein most similar to...

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Bibliographic Details
Published in:Fish & shellfish immunology 2000-01, Vol.10 (1), p.61-85
Main Authors: Harms, C.A, Kennedy-Stoskopf, S, Horne, W.A, Fuller, F.J, Tompkins, W.A.F
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Base Sequence
Bass - metabolism
Blotting, Northern - veterinary
Blotting, Southern - veterinary
Chimera - genetics
Cloning, Molecular
Consensus Sequence
Humans
Kidney - chemistry
Molecular Sequence Data
Morone chrysops
Morone saxatilis
Oncorhynchus mykiss
Rats
Reverse Transcriptase Polymerase Chain Reaction - methods
RNA, Messenger - analysis
Teleostei
Transforming Growth Factor beta - genetics
transforming growth factor- beta
transforming growth factor-β, TGF-β, sequence, hybrid striped bass, Morone saxatilis X M. chrysops, quantitative PCR
Xenopus
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Summary:A transforming growth factor (TGF)-β was isolated and cloned from hybrid striped bass (Morone saxatilis x M. chrysops) anterior kidney mononuclear cells. This isolate (Genbank accession number AF140363) contains an open reading frame of 1146 bases coding for a 382 amino acid protein most similar to rainbow trout TGF-β (57·3 and 78·6% identity with precursor and active protein, respectively) and rat TGF-β1(41·1 and 68·8% identity with precursor and active protein, respectively). Consensus primers were demonstrated to amplify specifically by polymerase chain reaction (PCR), a TGF-β segment from 14 species of teleost fish comprising 10 taxonomic families in 7 orders. A reverse transcription quantitative competitive polymerase chain reaction (RT-qcPCR) assay was devised to measure TGF-β mRNA expression in teleost fish. Higher levels of TGF-β mRNA expression were detected in mononuclear cells of peripheral blood than from spleen or anterior kidney.
ISSN:1050-4648
1095-9947
DOI:10.1006/fsim.1999.0230