Kumar R, Ghosh M, Kumar N, Balhara A K, Gupta M, Sharma R K, Singh I
Central Institute for Research on Buffaloes, Hisar, Haryana, India.
Department of Veterinary Biochemistry, Ranchi Veterinary College, Birsa Agricultural University, Kanke, Ranchi, Jharkhand, India.
Reprod Domest Anim. 2017 Jun;52(3):505-512. doi: 10.1111/rda.12941. Epub 2017 Mar 12.
The enormous production potential of buffaloes has never been accomplished due to various reproductive insufficiencies. Among them, post-partum anoestrus, a multifactorial disorder, is predominant but any genetic association is yet to be established. This study focused to identify novel polymorphisms in heat-shock protein 70 (HSP70) gene and its possible association with post-partum anoestrus in Murrah buffaloes. A 579-bp fragment from 5' untranslated region of HSP70 gene was amplified by polymerase chain reaction from blood genomic DNA of 614 animals maintained under similar management conditions. In phase-I experiment, custom sequencing and restriction enzyme (RE) digestion of the amplified fragment were performed in 40 buffaloes with similar post-partum oestrous conditions over previous consecutive three or more gestations-20 animals each showing post-partum anoestrus (>120 days after parturition) and normal cyclicity (<65 days after parturition). While in phase-II experiment, herd screening by RE analysis was performed in remaining 574 animals. Four transversions at T-75G, C+31G, T+38G and C+97A and three transition mutations at T-153C, T+33C and A+44G positions were observed. Polymorphism at T+38G site revealed significant (p < .05) variation, where homozygous G was present only in post-partum anoestrous animals while nucleotide T was present randomly in both groups of phase-I animals whereas phase-II experiments revealed homozygous G in 55 animals. Regression analysis in relation to average post-partum interval against genotypic frequencies at T+38G also depicted significant association. HSP70 gene polymorphism at T+38G position can therefore be used as genetic marker for excluding probable post-partum anoestrous buffaloes from herd for breeding programmes.
由于各种生殖功能不全，水牛巨大的生产潜力从未得到充分发挥。其中，产后乏情是一种多因素紊乱疾病，最为常见，但尚未确定其与任何基因的关联。本研究旨在鉴定热休克蛋白70（HSP70）基因中的新多态性及其与摩拉水牛产后乏情的可能关联。通过聚合酶链反应从614头处于相似管理条件下的动物血液基因组DNA中扩增出HSP70基因5'非翻译区的一个579 bp片段。在第一阶段实验中，对40头在过去连续三个或更多妊娠期具有相似产后发情情况的水牛进行了定制测序和扩增片段的限制性内切酶（RE）消化——每组20头，分别表现为产后乏情（分娩后>120天）和正常发情周期（分娩后<65天）。而在第二阶段实验中，对其余574头动物进行了RE分析群体筛查。观察到在T - 75G、C + 31G、T + 38G和C + 97A位置有四个颠换，在T - 153C、T + 33C和A + 44G位置有三个转换突变。T + 38G位点的多态性显示出显著（p <.05）差异，其中纯合G仅存在于产后乏情动物中，而核苷酸T在第一阶段实验的两组动物中随机分布，而第二阶段实验显示55头动物中有纯合G。T + 38G基因型频率与平均产后间隔的回归分析也显示出显著关联。因此，T + 38G位置的HSP70基因多态性可作为遗传标记，用于在育种计划中从牛群中排除可能产后乏情的水牛。
