首页|暗纹东方鲀TGF-β1基因的克隆、表达特征及生长性状相关SNP位点筛选

暗纹东方鲀TGF-β1基因的克隆、表达特征及生长性状相关SNP位点筛选

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为探究暗纹东方鲀(Takifugu fasciatus)TGF-β1基因与其生长性状的关联及作用,采用RACE技术克隆了暗纹东方鲀TGF-β1基因,分析了其基因结构与表达特征,并通过直接PCR扩增法对TGF-β1基因进行多片段比对,在此基础上筛选出与生长性状相关的SNP位点.研究发现TGF-β1基因cDNA全长共2 217 bp,包含5'UTR51 bp,3'UTR 1 014 bp,ORF 1 152 bp,编码383个氨基酸.氨基酸序列比对分析表明,暗纹东方鲀TGF-β1基因与红鳍东方鲀(Takifugu rubripes)同源性最高(92.82%).进化树分析表明,暗纹东方鲀TGF-β1基因编码氨基酸序列与红鳍东方鲀聚为一支,与鲈形目鱼类亲缘关系最近(68.46%~73.33%).qRT-PCR检测显示,TGF-β1基因在暗纹东方鲀8种不同组织(心脏、肝脏、脑、脾脏、肾脏、肌肉、中肠和鳃)都有表达,其中肾脏表达量最高,心脏和脾脏表达量次之.通过SNP位点不同基因型与个体生长性状的相关性分析,发现TGF-β1上的SNP g.3908 C>A位点与暗纹东方鲀多项生长性状有关,其中CC型是优势基因型,且CC型个体体宽、肥满度较高.研究结果证实TGF-β1基因与暗纹东方鲀生长性状有一定的相关性,可为暗纹东方鲀分子标记辅助育种提供基础资料.
Molecular cloning,expression analysis and growth traits-ralated SNP loci screening of TGF-β1 gene in Takifugu fasciatus
Takifugu fasciatus is an important farmed fish in China with high nutritional and economic value.The natural germplasm resources of T.fasciatus are getting depleted with the deterioration of living environment.Artificial breeding has become a necessary measure to alleviate the contradiction between supply and demand of T.fasciatus in the market and to slow down the decrease of wild resources of T.fasciatus in natural environment.In the absence of rejuvenation of wild resources and updating of genetic information of T.fasciatus,inbreeding occurred in the population.The degradation of germplasm is manifested in many aspects such as uneven growth,gradual decline of individual resistance and slow growth rate.TGF-β1,a multi-effect cytokine of the transforming growth factor β family,is related to cell proliferation and differentiation,cell proliferation inhibition,tissue formation,apoptosis regulation and immune regulation,and also plays an important role in the growth and development of fish.In order to investigate the association and role of TGF-β1 gene with growth traits in T.fasciatus,healthy T.fasciatus were selected to clone the full-length cDNA sequence of TGF-β1 gene by RACE technology.Further,to characterize the expression characteristics of TGF-β1 gene in 8 different tissues(heart,liver,brain,spleen,kidney,muscle,midgut and gill)by qRT-PCR.Finally,we selected 300 T.fasciatus at the same period.And then we measured their 10 quantitative characters and calculated one proportional character(condition factor).We also developed and analyzed the SNP loci on TGF-β1 of T.fasciatus by PCR direct sequencing and correlated the growth traits-related SNP loci screening of TGF-β1 gene in T.fasciatus by sequence multiplexed analysis.The results showed:1)the cDNA sequence of TGF-β1 was 2 217 bp,composing of 51 bp at the 5'end,1 014 bp at the 3'end and 1 152 bp at the ORF,encoding 383 amino acids.Amino acids sequence alignment analysis showed that TGF-β1 of T.fasciatus had high homology of 92.82%with T.rubripes.The phylogenetic tree analysis showed that T.fasciatus TGF-β1 clustered with T.rubripes.The full-length cDNA sequence of TGF-β1 gene of T.fasciatus was similar to that of most pelagic fish,and had the closest relationship with Perciformes and the farthest relationship with Gekko japonicus.2)TGF-β1 gene expression data showed that TGF-β1 gene was expressed in different levels in 8 tissues of T.fasciatus.The expression level of TGF-β1 was the highest in kidney,followed by heart and spleen,while the expression was lower in the other 5 tissues with no significant difference from each other(P<0.05).3)TGF-β1 gene SNP loci of T.fasciatus were developed,and a mutation in the nucleotide sequence at position 3908 in TGF-β1 was found,named SNP g.3908 C>A.After comparison,it was found that SNP g.3908 C>A was located on the intron of TGF-β1 in T.fasciatus,and did not participate in the transcription and translation of TGF-β1.Correlation analysis between different genotypes of SNP loci and growth traits showed that SNP g.3908 C>A on TGF-β1 was related to many growth traits of T.fasciatus,among which the CC type was the dominant genotype,and CC type individuals had higher body width and plumpness.The results confirm that TGF-β1 gene has a correlation with the growth traits of T.fasciatus,which is expected to provide reference materials for molecular marker-assisted breeding of T.fasciatus.

Takifugu fasciatusTGF-β1expression analysisgrowth traitsSNP

马思思、喻伟峰、张凯、张雯雯、唐晓东、刘雨曦、尹绍武、王涛

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南京师范大学海洋科学与工程学院,南京 210023

江苏省特色水产育种与绿色高效养殖技术工程研究中心,南京 210023

暗纹东方鲀 TGF-β1 表达特征 生长性状 SNP

江苏省种业振兴"揭榜挂帅"项目江苏省农业科技自主创新项目国家自然科学基金国家自然科学基金国家重点研发计划江苏省自然科学基金青年基金

JBGS2021034CX22202932172948318004362018YFD0900301BK20210563

2024

海洋渔业
中国水产学会 中国水产科学研究院东海水产研究所

海洋渔业

CSTPCD北大核心
影响因子:1.063
ISSN:1004-2490
年,卷(期):2024.46(2)
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