热休克蛋白70(HSP70)SacII多态性抑制剂对抗西方来源的局部耐热性

Kusnadidi Subekti, Dedi Duryadi Solihin, R. Afnan, A. Gunawan, Cece Sumanri
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引用次数: 1

摘要

摘要本研究的目的是确定热休克蛋白70(HSP70)基因多态性对耐热性的影响。对4个西苏门答腊鸭种群的110只当地鸭子(胶带、阴影、床和雨伞)进行了基因分型。所用的限制性内切酶是SacII。用24尾当地鸭子样本进行1小时的热注射,以分析血液学状况。PCR-RFLP和Sanger测序法用于鉴定多态性。使用MEGA7软件进行多态性分析,并使用SAS9.4软件进行ANOVA分析其影响。产生的扩增产物为466个碱基对。HSP70/SacII位点识别一个SNP位点(单核苷酸多态性)g.1702TC,产生两个等位基因(T和C)和三个基因型(TT、TC、CC)。HSP70/SacII基因在所有群体中都具有多态性。研究表明,在所有人群中,T等位基因的频率都高于C等位基因。卡方分析(χ2)表明,所有本地鸭种群都满足hardy-Weinberg平衡缺口。HSP70基因型的血液学指标显示异叶值和H/L值之间存在差异(P0.05),CT基因型表现出比其他基因型更好的耐热性。基于多样性及其对血液学状态的影响,HSP70/SacII基因可以作为本地鸭耐热性的分子标记。热休克蛋白70(HSP70)SacII基因多态性对西苏门答腊土鸭耐热性的影响本研究的目的是确定HSP70/SacII基因多态性对耐热性的影响。对来自西苏门答腊鸭群的110只本地鸭进行了基因分型。所用的限制性内切酶是SacII。对多达24只鸟的样本进行1小时的热应激,以分析血液学状况。PCR-RFLP和Sanger测序分析用于鉴定多态性。用MEGA7软件进行多态性分析,并用统计学SAS9.4软件进行方差分析。扩增产物为466bp。HSP70/SacII识别一个SNP g.1702TC,产生两个等位基因(T和C)和三种基因型(TT、TC、CC)。HSP70/SacII基因座在所有群体中均具有多态性。在所有人群中,T等位基因的频率均高于C等位基因。卡方分析(χ2)表明,所有地方鸭种群均符合Hardy-Weinberg平衡。HSP70/SacII基因对血液学状态的影响显示,CT基因型的异嗜性和H/L比差异(P0,05)比其他基因型表现出更好的耐热性。HSP70/SacII基因的多态性及其对血液学状态的影响可作为本地鸭耐热性的标志。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Pengaruh Polimorfisme Gen Heat Shock Protein 70 (HSP70) SacII terhadap Toleransi Panas Itik Lokal Sumatera Barat
ABSTRAK. Tujuan dari penelitian ini adalah untuk mengidentifikasi pengaruh polimorfisme gen heat shock protein 70 (HSP70) terhadap sifat toleransi panas. Genotiping dilakukan pada 110 itik lokal dari 4 populasi itik Sumatera Barat (pitalah, bayang, kamang, dan payakumbuh). Enzim restriksi yang digunakan adalah SacII. Cekaman panas dilakukan selama 1 jam dengan sampel itik lokal sebanyak 24 ekor untuk menganalisa status hematologi. PCR-RFLP dan Sanger sequencing assay digunakan untuk mengidentifikasi polimorfisme. Analisis polimorfisme dilakukan dengan menggunakan Software MEGA 7 dan pengaruhnya dianalisis dengan ANOVA menggunakan software SAS 9.4. Produk amplifikasi yang dihasilkan yaitu 466 pasang basa. Lokus HSP70/SacII mengenali satu situs SNP (Single nucleotide Polymorphism) g.1702TC, menghasilkan dua alel (T dan C) dan tiga genotype (TT, TC, CC). Gen HSP70/SacII bersifat polimorfik pada semua populasi. Hasil penelitian menunjukkan bahwa alel T memiliki frekuensi yang lebih tinggi daripada alel C pada semua populasi. Analisis chi-kuadrat (χ2) menunjukkan bahwa semua populasi itik lokal memenuhi kaidah kesetimbangan Hardy-Weinberg. Pengaruh genotipe gen HSP70 terhadap status hematologi menunjukkan perbedaan (P0,05) terhadap nilai heterofil dan nilai H/L dengan genotipe CT menunjukkan toleransi panas yang lebih baik dibanding genotipe lainnya. Gen HSP70/SacII dapat dijadikan marka molekuler sifat toleransi panas pada itik lokal berdasarkan keragaman dan pengaruhnya terhadap status hematologi.  (The effect of heat shock protein 70 (HSP70) SacII gene polymorphism on heat tolerance of West Sumatera local duck) ABSTRACT. The objective of this study was to identify the effect of HSP70/SacII gene polymorphism associated with the thermotolerance trait. Genotyping was performed on 110 local duck from the West Sumatera ducks population (pitalah, bayang, kamang, and payakumbuh). The restriction enzyme used was SacII. Heat stress was done for 1 hour with samples used as much as 24 birds to analyze hematological status. PCR-RFLP and Sanger sequencing assays were used to identify polymorphism. Analysis of polymorphism was conduct by MEGA 7 software and its effect is analyzed with ANOVA using Statistical SAS 9.4 software. The product of amplification was 466 bp. HSP70/SacII recognized one SNP g.1702TC, producing two alleles (T and C) and three genotypes (TT,TC,CC). The HSP70/SacII locus were polymorphic in all population. T allele had a higher frequency than C allele in all populations. The analysis of chi-square (χ2) showed that all local ducks population were fit with Hardy-Weinberg equilibrium. The effect of HSP70/SacII gene on hematological status showed differences (P0,05) of heterophil and H/L ratio with CT genotype show better heat tolerance than other genotypes. HSP70/SacII gene can be used as a marker of heat tolerance in local ducks based on polymorphism and its effect on hematological status.
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