黄颡鱼生长激素基因多态性与生长性状的关联性研究
本文关键词: 黄颡鱼 GH基因 单核苷酸多态性 微卫星 生长性状 出处:《华南农业大学》2016年硕士论文 论文类型:学位论文
【摘要】:本研究对黄颡鱼生长激素基因全长进行克隆测序,分析GH在组织中的差异性表达,对GH基因的单核苷酸多态性(SNP)和微卫星(SSR)多态性与主要的生长性状进行关联分析,并获得调控经济性状的优势基因位点。(1)GH基因的分子特征根据亲缘关系较近物种的GH基因的保守区域设计引物,通过克隆测序拼接后获得DNA全长序列,共3117bp,包括5′UTR 165bp,3′UTR 485bp,由5个外显子和4个内含子组成,外显子大小分别为10bp、140bp、117bp、132bp、204bp,内含子大小分别为302bp、85bp、642bp、835bp。(2)GH基因的差异性表达通过半荧光定量和实时荧光定量两种方法研究黄颡鱼GH基因mRNA的差异性表达。结果显示:GH基因只在垂体中特异表达,而在肌肉、脾脏、脂肪、肾脏、胃、盲肠、心脏、头肾、精巢、肠、鳃、肝脏、鱼皮和脑这14个组织中均未检测到表达。(3)SNP与生长性状的关联性以GH基因作为影响黄颡鱼主要生长性状的候选基因,样本数为160,通过PCR产物直接测序法筛选SNP位点,筛选区域为片段外显子4到片段外显子5和3′UTR区域,共检测到5个SNP位点,内含子4区域检测到4个,3′UTR发现1个。并对SNP位点基因型和主要生长性状(体重、全长、体长、体高、体厚、头长、尾柄长、尾柄高)进行关联分析,结果显示:存在于内含子4的位点1674GA和位点2154TG与生长性状无显著关联;位点2100AG与性状体厚有极显著关联(P0.01),GG为优势基因型;位点2285TC与尾柄长有显著关联(P0.05),CC为优势基因型;存在于3′UTR的位点2822AG与全长、尾柄高有显著关联(P0.05),与体长、尾柄长有极显著关联(P0.01),GG为优势基因型。(4)SSR与生长性状的关联性通过SSRHunter软件在黄颡鱼GH基因上查找到4个微卫星位点,将其在96-毛细管ABI3730xl上进行检测,根据检测结果判定基因型,并与生长性状进行关联分析。结果表明:位点AT与生长性状无显著关联;位点GA与体重、体长、体高、体厚有显著关联(P0.05),与全长、尾柄高有极显著关联(P0.01),CD为优势基因型;位点TCTT与头长、尾柄高有显著关联(P0.05),BD为优势基因型;位点AC与体厚、头长有显著关联(P0.05),DD为优势基因型。
[Abstract]:In this study, the full-length growth hormone gene of Pelteobagrus fulvidraco was cloned and sequenced, and the differential expression of GH in tissues was analyzed. Single nucleotide polymorphisms (SNPs) and microsatellite SSRs (SSRs) of GH gene were associated with major growth traits. The molecular characteristics of the GH gene were designed according to the conserved region of the GH gene of the nearest species. The full-length sequence of DNA was obtained by cloning and sequencing, which consisted of 5 exons and 4 introns. The size of exon is 10bp, 140bp, 117bp, 132bp, 204bp, and the intron is 302bp, 85bp, 642bp, respectively. 835bp. / 2). The differential expression of GH gene in Pelteobagrus fulvidraco was studied by semi-fluorescent quantitative method and real-time quantitative method. The results showed that the specific expression of the GH gene was found only in pituitary gland of Pelteobagrus fulvidraco. But in muscle, spleen, fat, kidney, stomach, cecum, heart, head kidney, testis, intestine, gills, liver. There was no correlation between the expression of SNP and growth traits in fish skin and brain. The GH gene was used as the candidate gene to affect the main growth traits of Pelteobagrus fulvidraco. The sample number was 160. The SNP loci were screened by direct sequencing of PCR products. Five SNP loci were detected in the regions from exon 4 to exon 5 and exon 3. SNP locus genotypes and main growth traits (body weight, full length, body length, body height, body thickness, head length, tail stalk length) were detected in intron 4 region. The results showed that there was no significant correlation between locus 1674GA and locus 2154TG in intron 4. Locus 2100AG was significantly associated with the body thickness of the trait P0.01GG was the dominant genotype. Locus 2285TC was significantly associated with the length of caudal stalks. P0.05C was the dominant genotype. There was a significant correlation between the height of the tail stipe and the body length, the length of the tail stalk and the length of the tail stipe (P0.01). Four microsatellite loci were found on the GH gene of Pelteobagrus fulvidraco by SSRHunter software. It was detected on 96- capillary ABI3730xl, and genotypes were identified according to the detection results. The results showed that there was no significant correlation between AT locus and growth traits. There was a significant correlation between GA and body weight, body length, body height, body thickness, and total length. There was a significant correlation between TCTT locus and head length and height of caudal stalks (P0.05BD) as dominant genotypes. There was a significant correlation between AC and body thickness and head length.
【学位授予单位】:华南农业大学
【学位级别】:硕士
【学位授予年份】:2016
【分类号】:S917.4
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