Nucleotide sequence and mRNA expression analysis of <Emphasis Type="Italic">β</Emphasis>-actin gene in the orange-spotted grouper <Emphasis Type="Italic">Epinephelus coioides</Emphasis> |
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Authors: | Email author" target="_blank">W?M?ZhangEmail author Y?Zhang L?H?Zhang S?G?Wang T?Y?Zhu D?Lin G?Z?Ma |
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Institution: | (1) Institute of Aquatic Economic Animals and Guangdong Provincial Key Laboratory for Aquatic Economic Animals, College of Life Sciences, Zhongshan (Sun Yat-Sen) University, Guangzhou, 510275, P.R. China;(2) School of Life Sciences, Normal University of Southern China, Guangzhou, 510631, P.R. China |
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Abstract: | The full-length cDNA, encoding the orange-spotted grouper β-actin and spanning 1920 bp including a poly (A) tail, was cloned
from its brain cDNA library. The open reading frame encodes a protein of 375 amino acids. Sequence analysis indicated that
it contained the typical structural features of cytoplasmic actins, and showed higher homology with other vertebrate β-actin
than any other members of the actin family. The partial genomic sequence indicated that the organization of the β-actin gene
in the orange-spotted grouper might also be conserved. Northern blot analysis indicated that it was expressed at high levels
in the brain, spleen, adipose tissue, ovary, and liver, but at low levels in the gill filament and heart, and at a very low
level in the kidney. The expression of β-actin gene in the skeletal muscle was barely detectable. These results indicated
that the expression of the orange-spotted grouper β-actin gene showed significant variation in different tissues. Therefore,
caution should be taken when using β-actin gene as an internal control in the normalization of gene expression among tissues.
Whereas, semi-quantitative RT-PCR analysis indicated that treatment with 17α–methyltestosterone (MT) had little effect on
the mRNA expression of β-actin gene in the in vitro incubated hypothalamus, pituitary, and ovary fragments of the orange-spotted grouper, suggesting β-actin can be used as an
internal control for RT-PCR analysis of MT effects on gene expression in these tissues. |
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Keywords: | β -actin cDNA internal control 17α – methyltestosterone mRNA expression orange-spotted grouper (Epinephelus coioides) |
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