罗华伦, 张依裕, 李万贵, 王单单, 余蓉蓉. 2017: 鸭SCD1基因启动子多态性对血清生化指标的遗传效应. 南方农业学报, 48(10): 1891-1898. DOI: 10.3969/j.issn.2095-1191.2017.10.25
引用本文: 罗华伦, 张依裕, 李万贵, 王单单, 余蓉蓉. 2017: 鸭SCD1基因启动子多态性对血清生化指标的遗传效应. 南方农业学报, 48(10): 1891-1898. DOI: 10.3969/j.issn.2095-1191.2017.10.25
LUO Hua-lun, ZHANG Yi-yu, LI Wan-gui, WANG Dan-dan, YU Rong-rong. 2017: Genetic effects of SCD1 gene promoter polymorphism on serum biochemical indexes in ducks. Journal of Southern Agriculture, 48(10): 1891-1898. DOI: 10.3969/j.issn.2095-1191.2017.10.25
Citation: LUO Hua-lun, ZHANG Yi-yu, LI Wan-gui, WANG Dan-dan, YU Rong-rong. 2017: Genetic effects of SCD1 gene promoter polymorphism on serum biochemical indexes in ducks. Journal of Southern Agriculture, 48(10): 1891-1898. DOI: 10.3969/j.issn.2095-1191.2017.10.25

鸭SCD1基因启动子多态性对血清生化指标的遗传效应

Genetic effects of SCD1 gene promoter polymorphism on serum biochemical indexes in ducks

  • 摘要: 目的分析鸭SCD1基因启动子多态性与血清生化指标的遗传效应,揭示鸭SCD1基因启动子的生理调控机制,为今后开展畜禽SCD1基因研究及遗传标记选育提供参考依据.方法构建基因组DNA池,利用PCR-RFLP结合DNA直接测序技术检测鸭SCD1基因启动子区遗传变异情况,采用MegAlign寻找SNP位点,通过在线启动子分析软件预测SNP位点对鸭SCD1基因启动子g.952260~g.954527区域转录因子的影响,并以SPSS 19.0分析启动子酶切位点变异对父母代樱桃谷鸭血清生化指标的遗传效应.结果从樱桃谷鸭SCD1基因启动子g.952260~g.954527区域共检测到11个SNPs位点.其中,g.952323C>A和g.952661A>G位点突变分别致使原有转录因子D1和Sp1消失;g.952702A>G位点突变则产生新的转录因子TEC1;g.952591T>C、g.952868A>T、g.952869T>G、g.952971G>C和g.953301T>C突变区域无转录因子结合位点,也未产生新的转录因子结合位点;g.952873T>G和g.954401T>C位点突变未引起转录因子改变;g.954239C>T位点突变导致原有转录因子Sp1、Sp1、Tra-1和AP-2α改变为Sp1、Tra-1、ETF和Sp1.通过PCR-Bgl II-RFLP和直接测序比对分析,发现g.952868A>T和g.952869T>G双碱基突变造成原始序列AGT952868G952869CT突变成AGA952868T952869CT,而产生新的Bgl II酶切位点,共产生3种基因型:CC(2268 bp)、CD(2268+1659+609 bp)和DD(1659+609 bp),2个等位基因(C和D).CC基因型和C等位基因分别为优势基因型和优势等位基因,其频率分别为0.710和0.805;有效等位基因数(Ne)和多态信息含量(PIC)分别为1.458和0.265,属中度多态位点;卡方(χ2)检验结果表明,Bgl II酶切位点突变所产生的基因型分布严重偏离哈代—温伯格平衡(χ2>χ20.01,P<0.01).Bgl II酶切位点变异与父母代樱桃谷鸭血清生化指标的关联性分析结果表明,总体上以DD基因型樱桃谷鸭的血清生化指标略优于其他两种基因型.结论樱桃谷鸭SCD1基因启动子g.952260~g.954527区域共筛查到11个SNPs位点,其中g.952868A>T和g.952869T>G位点变异产生新的Bgl II酶切位点,且该酶切位点变异可能对鸭血清生化指标有调控效应.

     

    Abstract: ObjectiveThe genetic effects of duck SCD1 gene promoter polymorphism and serum biochemical indexes were analyzed to reveal the physiological regulation mechanism of duck SCD1 gene promoter and provide reference for future research on gene SCD1 and breeding of genetic markers.MethodGenetic variation of duck SCD1 gene promoter region was detected through a combination use of polymerase chain reaction restriction and fragment length polymorphism (PCR-RFLP)and DNA direct sequencing technology on the basis of constructing a DNA pool. Single nucleotide polymor-phisms(SNP)loci were aligned by online promoter analysis software to predict the effects of SNP loci on transcription factors in duck SCD1 gene promoter region(g.952260-g.954527). Genetic effects of promoter restriction site variation on serum biochemical indexes in parental generation Cherry Valley ducks were analyzed by SPSS 19.0 software.ResultEleven SNPs loci were detected in g.952260-g.954527 region in Cherry Valley duck SCD1 gene promoter. Among them, g. 952323C>A and g. 952661A>G site mutations led to disappearance of the original transcription factors D1 and Sp1 re-spectively;g. 952702 A>G site mutation produced a new transcription factor TEC1;no transcription factor binding sites or new transcription factor binding sites were found in g. 952591T>C,g. 952868A>T,g. 952869T>G,g. 952971G>C and g. 953301T>C mutation regions;g. 952873T>G and g. 954401T>C mutations led no change to transcription factors;g.954239C>T site mutation changed original transcription factors from Sp1,Sp1,Tra-1,and AP-2αto Sp1,Tra-1,ETF, and Sp1. Comparative analysis by PCR-Bgl II-RFLP method and direct sequencing showed that g. 952868A>T and g. 952869T>G double base mutation led to the change of original sequence from AGA952868T952869CT to AGT952868G952869CT, which produced new Bgl II enzyme cutting site. Three genotypes namely CC(2268 bp),CD(2268+1659+2268 bp)and DD(1659+609 bp),and two alleles(C and D)were produced. CC genotype and C allele were dominant genotype and dominant allele respectively,whose frequencies were 0.710 and 0.805. Effective alleles(Ne)and polymorphism informa-tion content(PIC) were 1.458 and 0.265 respectively,belonging to moderate polymorphism. Chi-square(χ2) results showed that genotypes distribution caused by Bgl II enzyme cutting site mutation greatly deviated from the Hardy-Wein-berg equilibrium(χ2>χ20.01,P<0.01). The correlation analysis between Bgl II enzyme cutting site mutation and serum bio-chemical indexes in parental Cherry Valley ducks showed that the serum biochemical markers of DD genotype in Cherry Valley duck were slightly better than those of the other two genotypes.ConclusionEleven SNPs loci are detected in SCD1 gene promoter region g. 952260-g. 954527 of Cherry Valley duck. Among them,g. 952868A>T and g. 952869T>G site variations produce a new Bgl II enzyme cutting site,and this enzyme cutting site may have regulatory effects on duck serum biochemical indexes.

     

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