联系我们 加入收藏
 
年期检索 高级检索
33
2025年4月6日 星期日
  2017, Vol. 25 Issue (4): 599-606    
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
从江香猪LPL基因克隆、多态及时序表达图谱构建
张雄1,张勇2,尚以顺3,陈祥4,史开志3,白优1,陆静5
1. 贵州大学
2. 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
3. 贵州省畜牧兽医研究所
4. 贵州大学 动物科学学院/高原山地动物遗传育种与繁殖教育部重点实验室/贵州省动物遗传育种与繁殖重点实验室
5. 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
Cloning, Polymorphisms and Tissue Expression Profile of LPL Gene in Congjiang Pigs (Sus scrofa)
1, 1, 1, 1, 1, 1
全文: PDF (1253 KB)   HTML (1 KB) 
输出: BibTeX | EndNote (RIS)      
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(GenBank 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qRT-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(P<0.05),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(P>0.05)。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(P<0.05)。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。
服务
把本文推荐给朋友
加入我的书架
加入引用管理器
E-mail Alert
RSS
作者相关文章
张雄
张勇
尚以顺
陈祥
史开志
白优
陆静
关键词 从江香猪脂蛋白脂酶(LPL)克隆时序表达限制性长度多态性聚合酶链反应(PCR-RFLP)    
Abstract:Lipoprotein lipase (LPL) has the function of regulating fat deposition in animals, but its molecular genetic mechanisms is seldomly studied in local mini-pig (Sus scrofa) breeds. In order to understand the function of gene LPL and reveal its effects on meat quality traits in Congjiang pigs. In this study, the CDS region of LPL gene was cloned by RT-PCR from Congjiang pigs, the physical and chemical properties as well as structure characteristics of LPL protein was predicted using bioinformatics softwares. The genotype of AfaⅠlocus of intron 5 in Congjiang pigs was tested by polymerase chain reaction-restriction fragment length polymorphism technique (PCR-RFLP). Meanwhile, the expression profile of LPL gene was investigated by qRT-PCR and their association with content of intramuscular fat and activities of LPL in dorsal muscles of different stages Congjiang pigs were analyzed. The results showed that the CDS of LPL gene was 1 437 bp encoding a stable, hydrophilic protein with 478 amino acids and a PLAT/LH2 domain. There was high amino acid homology of LPL among different breeds of pig, the amino acid sequence of LPL of Congjiang pigs had similarity of 99.8% with Norwegian Landrace and Luchuan pigs, the lowest homology with Guizhou baixiang pigs was 99.2%. PCR-RFLP results showed that the AfaⅠlocus of the LPL gene have 3 genotypes in Congjiang pigs, i.e. CC, CG and GG, and the CG was dominant genotype. The allele frequencies of C and G at AfaⅠlocus were 0.350 5 and 0.649 5, repectively. The Congjiang pig population investigated was at Hardy-Weinberg equilibrium in this polymorphic sites. The skin thickness of CC genotype was significantly higher than that of CG genotype at AfaⅠlocus in Congjiang pigs (P<0.05). Furthermore, the backfat thickness, lion eye area and intramuscular fat content of CC were also higer than those of other genotypes (P>0.05). qRT-PCR results suggested that the expression level of LPL gene in the dorsal muscles of Congjiang pigs at different stages was decreased first and then increased, the correlation analysis indicated that there was a significant correlation between LPL mRNA expression and intramuscular fat content (P<0.05). In conclusion, the LPL gene might play an important role in the IMF deposition of Congjiang pigs, this study could provide a theoretical basis for the follow-up analysis of structure and function of LPL gene in Congjiang pigs.
Key wordsCongjiang pigs    Lipoprotein lipase (LPL)    Cloning    Temporal expression    Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)
收稿日期: 2016-11-21      出版日期: 2017-03-31
ZTFLH:  S828  
基金资助:贵州省科研机构服务企业行动计划项目(“生猪健康养殖配套技术服务企业行动计划”);贵州省工程中心建设项目;贵州大学引进人才科研项目(“提高香猪产仔数及仔猪成活率关键技术研究与应用”)
通讯作者: 张勇     E-mail: as.yzhang@gzu.edu.cn
引用本文:   
张雄 张勇 尚以顺 陈祥 史开志 白优 陆静. 从江香猪LPL基因克隆、多态及时序表达图谱构建[J]. , 2017, 25(4): 599-606.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2017/V25/I4/599
 
版权所有 © 2014 《农业生物技术学报》编辑部   京ICP备11035905号-3
地址:北京市海淀区圆明园西路2号中国农业大学生命科学楼1053室 邮编:100193
电话:010-62733684 传真:010-62731615 E-mail: nsjxb@cau.edu.cn