从江香猪LPL基因克

发布时间：2018-05-08 23:11

本文选题：从江香猪 + 脂蛋白脂酶(LPL)　；参考：《农业生物技术学报》2017年04期

【摘要】：脂蛋白脂酶(lipoprotein lipase,LPL)具有调控动物机体脂肪沉积的功能,但在小型地方猪种中LPL基因分子遗传机制研究鲜有报道。为探究LPL基因调控功能,揭示其对从江香猪(Sus scrofa)肉质性状的影响,本研究以从江香猪为实验动物,利用RT-PCR技术克隆从江香猪LPL基因CDS区(Gen Bank No.1994531),生物信息学软件预测蛋白理化性质及结构特征,通过限制性片段长度多态性聚合酶链反应(polymerase chain reaction-restriction fragment length polymorphism,PCR-RFLP)技术对从江香猪LPL基因第5内含子AfaⅠ位点进行遗传效应分析,运用q RT-PCR技术分析不同月龄阶段从江香猪背最长肌组织中LPL mRNA表达发育规律,并研究其与肌内脂肪(intramuscular fat,IMF)含量相关性。结果表明,从江香猪LPL基因CDS区全序列长1 437 bp,共编码478个氨基酸,构成具有PLAT/LH2结构域的稳定亲水性蛋白。不同品种猪间氨基酸序列的同源性很高,其中从江香猪与挪威长白猪和陆川猪氨基酸序列的同源性高达99.8%,与贵州白香猪同源性最低,为99.2%。PCR-RFLP分析结果显示,AfaⅠ位点共检测到CC、CG和GG 3种基因型,其中CG为优势基因型,等位基因C和G的频率分别为35.05%和64.95%,从江香猪群体在此位点处于Hardy-Weinberg平衡状态;CC基因型的皮厚显著高于CG型(P0.05),但背膘厚、眼肌面积和肌内脂肪含量与其他基因型个体间差异不显著(P0.05)。时序表达谱显示,不同月龄阶段从江香猪背最长肌组织中LPL mRNA表达水平呈现出先降低后升高的趋势,LPL mRNA表达量与IMF呈显著正相关(P0.05)。结果表明,LPL基因可作为影响从江香猪脂肪沉积的关键候选基因之一,实验结果可为从江香猪LPL基因结构功能的后续研究提供理论依据。
[Abstract]:Lipoprotein lipase lipase has the function of regulating fat deposition in animal body, but there are few reports on the molecular genetic mechanism of LPL gene in small local pig breeds. In order to investigate the regulatory function of LPL gene and reveal its effect on meat quality traits of Congjiang Xiang Sus scrofa, Congjiang Xiang pig was used as experimental animal in this study. Gen No. 1994531 was cloned from the CDS region of Congjiang Xiang pig LPL gene by RT-PCR technique. The bioinformatics software was used to predict the physicochemical properties and structural characteristics of the protein. Restriction fragment length polymorphism polymerase chain reaction-restriction fragment length polymorphism RFLP was used to analyze the genetic effect of Afa I locus in intron 5 of LPL gene in Congjiang Xiang pig. Q RT-PCR technique was used to analyze the expression and development of LPL mRNA in the longissimus dorsi muscle tissues of Congjiang Xiang pigs at different ages, and the correlation between LPL mRNA expression and intramuscular fat content was studied. The results showed that the complete CDS region of Congjiang Xiang pig LPL gene was 1 437 BP long, encoding 478 amino acids, and a stable hydrophilic protein with PLAT/LH2 domain was constructed. The homology of amino acid sequence among different breeds of pigs was very high. The amino acid sequence of Congjiang Xiang pig was 99.8 with Norway Landrace pig and Luchuan pig, and the lowest homology with Guizhou white Xiang pig. The results of 99.2%.PCR-RFLP analysis showed that there were three genotypes of CCG and GG, among which CG was the dominant genotype. The frequencies of alleles C and G were 35.05% and 64.95%, respectively. The skin thickness of CC genotype was significantly higher than that of CG-type P0.05, but the backfat thickness was significantly higher than that of CG-type P0.05G. There was no significant difference in eye muscle area and intramuscular fat content among individuals with other genotypes (P 0.05). The expression of LPL mRNA in the longissimus dorsi muscle of Conjiangxiang pig showed a tendency of decreasing at first and then increasing. The expression of LPL mRNA was positively correlated with IMF (P 0.05). The results showed that the LPL gene could be used as one of the key candidate genes affecting fat deposition in Congjiang Xiang pig, and the results could provide theoretical basis for further study on the structure and function of LPL gene in Congjiang Xiang pig.
【作者单位】：贵州大学高原山地动物遗传育种与繁殖省部共建教育部重点实验室/贵州省动物遗传育种与繁殖重点实验室;贵州省畜牧兽医研究所;
【基金】：贵州大学引进人才科研项目(贵大人基合字[2013]21号) 贵州省科研机构服务企业行动计划项目(黔科合服企[2015]4003号) 贵州省工程中心建设项目(黔科合农C字[2011]4022号)
【分类号】：S828

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