Molecular cloning and sequence analysis of a cDNA encoding pituitary thyroid stimulating hormone -subunit of the Chinese soft-shell turtle Pelodiscus sinensis and regulation of its gene expression
Resource
General and Comparative Endocrinology 146 (2006) 74–82
Journal
General and Comparative Endocrinology
Journal Volume
146
Journal Issue
2006
Pages
-
Date Issued
2006
Date
2006
Author(s)
Lin, Yao-Sung
Chien, Jung-Tsun
Chowdhury, Indrajit
Liao, Ching-Fong
Shen, San-Tai
Yu, Yuh-Lin John
DOI
246246/2006111501275261
Abstract
A cDNA encoding thyroid stimulating hormone -subunit (TSH ) was cloned from pituitary of the Chinese soft-shell turtle, Pelodiscus
sinensis, and its regulation of mRNA expression was investigated for the Wrst time in reptile. The Chinese soft-shell turtle TSH
cDNA was cloned from pituitary RNA by reverse transcription and polymerase chain reaction (RT-PCR), and rapid ampliWcation
cDNA end (RACE) methods. The Chinese soft-shell turtle TSH cDNA consists of 580-bp nucleotides, including 67-bp nucleotides of 5 -
untranslated region (UTR), 402-bp of the open reading frame, and 97-bp of 3 -UTR followed by a 14 poly (A) trait. It encodes a precursor
protein molecule of 133 amino acids with a putative signal peptide of 19 amino acids and a putative mature protein of 114 amino
acids. The number and position of 12 cysteine residues, presumably forming six disulWde bonds, one putative asparagine-linked glycosylation
site, and six proline residues that are found at positions for changing the backbone direction of the protein have been conserved in
the turtle as in other vertebrate groups. The deduced amino acid sequence of the Chinese soft-shell turtle TSH mature protein shares
identities of 82–83% with birds, 71–72% with mammals, 49–57% with amphibians, and 44–61% with Wsh. The Chinese soft-shell turtle
pituitaries were incubated in vitro with synthetic TRH (TSH-releasing hormone), thyroxine and triiodothyronine at doses of 10¡10 and
10¡8 M. TRH stimulated, while thyroid hormones suppressed, TSH mRNA levels in dose-related manner. The sequences of cDNA and
its deduced peptide of TSH as well as the regulation of its mRNA level were reported for the Wrst time in reptile.
Subjects
floorplanning
physical design
Chinese soft-shell turtle
Pituitary
Thyrotropin (TSH ) subunit
cDNA
Amino acid sequence
mRNA
TRH
Thyroid hormone
Publisher
Taipei:National Taiwan University Grad Inst Elect Engn
Type
other
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