가지마목의 stanniocalcin-1 (STC-1)의 유전자 분석과 환경수의 염분이 STC-1 mRNA 발현에 미치는 영향
Molecular characterization of stanniocalcin-1 (STC-1) and effect of environmental salinities on the mRNA in pleuronectiformes
- 발행기관 강릉대학교 일반대학원
- 지도교수 손영창
- 발행년도 2007
- 학위수여년월 2007. 2
- 학위명 석사
- 학과 및 전공 해양생명공학과
- 원문페이지 iv, 30 p.
- 본문언어 한국어
초록/요약
Stanniocalcin-1 (STC-1) is an anti-hypercalcemic hormone that is produced by the corpuscles of Stannius (CS) in the teleost. The hormone is a homodimeric glycoprotein involved in the calcium and phosphate regulation in both teleost fish and mammals. In the present study, I have cloned and characterized a full-length cDNA of STC-1 from the CS in turbot (Scophthalmus maximus) and olive flounder (Paralichyhus olivaceus). In addition, I examined its expression pattern in various tissues and in several tissues in different salinities. The turbot STC-1 cDNA (1246 nucleotides) encodes a preprohormone of 248 amino acids (aa) with a signal peptide of 18 aa and a pro-sequence peptide of 14 aa followed by a mature protein of 216 aa. Olive flounder STC-1 cDNA (1331 nucleotides) encoded a preprohormone of 251 aa with a signal peptide of 17 aa and a pro-sequence peptide of 14 aa followed by mature protein of 220 aa. RT-PCR analysis revealed that the turbot STC-1 gene is expressed in the CS, pituitary, brain, kidney, liver, heart, muscle, and ginad. In a transient transfection of turbot STC-1 cDNA construct into human embryonic kidney 293T cells followed by Western blot analysis, a recombinant STC-1 secreted into a medium was detected as a single band of 28 kDa under reducing condition and a 50 kDa protein under non-reducing condition. Deglycosylation of the recombinant STC-1 led to a decrease in the molecular mass. When juvenile olive flounder were adapted to 5, 15, 25 and 32 ppt salinities for 3 days, the STC-1 mRNA levels in the CS, gills, kidney, and intestine showed a decreasing pattern in accordance with freshwater ratio. The present study suggests that STC-1 gene is widely expressed and the secreted protein exists as a homodimeric glycoprotein. The STC-1 mRNA expression in the peripheral tissues and organs participating in ion-regulatory function is down-regulated by lower salinity in euryhaline fish species.
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