虎皮鱼PtScd1基因序列、功能与表达分析

doi:10.16378/j.cnki.1003-1111.2020.06.005

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Abstract Tiger skin barb Puntius tetrazona with body length of 5—7 cm was housed in a 30 L breeding tank at a rate of 30 fish per tank at water temperature of 27 ℃(control group), and then exposed to water temperature of 23 ℃, 19 ℃, 15 ℃, and 13 ℃ on the fourth day, by dropping 4 ℃ and 2 ℃ every 24 hours from the next day in the 4 groups. After 24 hours of treatment, 3 fish were randomly sampled from each tank, and anesthetized by MS-222 solution, and the brain, gills, liver and muscle tissues were dissected, and were quickly frozen in liquid nitrogen. Using CODEHOP software to design degenerate primers, a conserved sequence of tiger skin fish Scd1 (PtScd1) gene was amplified by PCR, RACE cloning and sequence splicing to obtain tiger skin fish stearoyl-CoA desaturase (SCD) PtScd1 gene cDNA sequence. A 1338 bp PtScd1 cDNA sequence was obtained by rapid-amplification of cDNA ends and sequence assembly, and the open reading frame was predicted with 981 bp, encoding 326 amino acids. PtSCD1 protein was predicted with a fatty-acid desaturase conservation domain according to the Conserved Domain Database. Swiss-Model showed that there was a highly conserved binding site for two zinc ions, with more likely to be a diiron center, in the dimensional structure of PtSCD1, with two unconserved ligand sites. The protein model indicated that PtSCD1 catalyzed the synthesis of monounsaturated fatty acids leading to introducing a double band into the delta-9 position of fatty acids chain. The real time quantitative PCR revealed that the descending abundance of PtScd1 transcripts in various tissues and organs of adult tiger barb fish was described as liver >brain >muscle >gill, with negative correlation between the relative expression level of PtScd1 transcripts in brain tissue and the exposure temperature, and tissue-specific expression of PtScd1 transcripts in a temperature-dependent way. The findings indicated that PtScd1 gene was expected to play a vital role for tiger skin barb coping with cold stress, which would help promote the research of tiger skin barb survival under cold temperature.

Key words： tropical stenothermal fish cold-tolerant gene PtScd1 protein structure prediction tissue-specific expression

Received: 24 September 2019

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	LIU Lili
	ZHANG Rong
	WANG Xiaowen
	ZHU Hua

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LIU Lili,ZHANG Rong,WANG Xiaowen, et al. Analysis of Sequence, Function and Expression of PtScd1 Gene in Tropical Stenothermal Fish Tiger Skin Barb Puntius tetrazona[J]. Fisheries Science, 2020, 39(6): 820-828.

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https://www.shchkx.com/EN/10.16378/j.cnki.1003-1111.2020.06.005 OR https://www.shchkx.com/EN/Y2020/V39/I6/820

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