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IMPDH基因克隆、生物信息学分析及在牦牛卵巢不同时期中的差异表达
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引用本文:熊显荣,兰道亮,胡嘉嘉,李 键,字向东.IMPDH基因克隆、生物信息学分析及在牦牛卵巢不同时期中的差异表达[J].西北农业学报,2017,26(6):812~819
DOI:10.7606/j.issn.1004-1389.2017.06.002
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作者单位
熊显荣,兰道亮,胡嘉嘉,李 键,字向东 (1.西南民族大学 生命科学与技术学院成都 610041 2.西南民族大学 青藏高原研究院成都 610041) 
基金项目:四川省科技支撑计划(2014NZ0114);西南民族大学中央高校基本科研业务费专项资金(2016NZYQN38)。
中文摘要:旨在获得牦牛IMPDH基因序列并进行生物信息学分析,同时分析其在卵巢组织中的时序表达谱,为进一步研究卵巢生长发育及排卵机制奠定基础。通过RT-PCR技术克隆牦牛的IMPDH基因,并运用生物信息学方法分析不同物种序列进化关系,利用荧光定量qRT-PCR技术检测IMPDH基因在不同发育时期牦牛卵巢组织中的表达情况。结果显示,克隆得到牦牛1 623 bp 的IMPDH基因cDNA 序列,内含1 545 bp开放阅读框序列,编码514个氨基酸残基。与野牦牛的相应序列具有很高的同源性(99.4%),表明IMPDH基因在进化过程中相对保守。实时定量RT-PCR检测结果显示IMPDH基因的表达随年龄的增加而呈现上升趋势。荧光定量qRT-PCR检测结果显示IMPDH基因在卵巢卵泡期表达最强,显著高于红体形成期。结果表明IMPDH参与牦牛卵巢组织的生长发育,并在卵巢活动中有明显的表达规律,表明IMPDH基因在牦牛繁殖周期具有重要作用,为进一步研究牦牛IMPDH基因功能奠定基础。
中文关键词:牦牛  克隆  IMPDH  序列分析  差异表达
 
Cloning, Bioinformatics Analysis and Differential Expression of IMPDH Gene in Yak Ovary at Different Stages
Abstract:The aim of this study is to obtain the information about sequence structure and bioinformatics analysis of yak IMPDH gene, analysis of sequential expression in ovarian tissue. It could provide information for further study of mechanism of ovarian development and ovulation. Yak IMPDH cDNA was cloned using reverse transcription polymerase chain reaction (RT-PCR), the sequences characteristics were analyzed, and IMPDH gene expression in ovary were examined by real time fluorescence quantitative PCR (qRT-PCR) at different stages of development and reproductive cycle. The study obtained a complete cDNA sequences of 1 623 bp of IMPDH gene of yak with a deduced protein, which were consisted of 514 amino acids that are highly homologous with other species.The sequence similarity of IMPDH between yak and wild yak was 99.4%, this indicated that IMPDH gene was conservative during evolution. The result of RT-PCR indicated that the expression of yak IMPDH increased with the increase of age. The IMPDH gene in ovarian follicular phase was the strongest expression at other stages, and significantly higher than that of the red body phase.These results suggested that IMPDH gene was probably related to the ovarian development, and there was a significantly expressed pattern during ovarian activity. These indicated that IMPDH played an important role during the reproductive cycle, and made a foundation for the further study of the function of IMPDH gene in yak.
keywords:Yak  Cloning   IMPDH  Sequence analysis  Differential expression
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