摘要
旨在揭示DLK1-DIO3印记域上一个新的长链非编码RNA(long noncoding RNA,lncRNA)LINC24065在自然繁殖牛(natural reproduction,NR)与体细胞核移植牛(somatic cell nuclear transfer,SCNT)中的组织表达与印记状态,对进一步了解DLK1-DIO3印记域在供体核重编程中的作用提供一定的理论基础。本研究以自然繁殖牛的大脑组织为试验材料,利用RACE和RT-PCR技术,在牛的DLK1-DIO3印记域内鉴定了一个新的印记的lncRNA基因,命名为LINC24065。用基于SNP(single nucleotide polymopisms)的RT-PCR产物直接测序方法分析LINC24065在自然繁殖牛和体细胞核移植牛组织中的表达及印记状态。序列分析发现,LINC24065编码6个可变剪接体,并在自然繁殖牛被检测的7个组织中都表达,而在体细胞核移植牛组织中没有检测到可变剪接体LINC24065-V3,且其它5个剪接体呈现组织特异性表达。印记状态分析发现,LINC24065在自然繁殖牛和体细胞核移植牛的7个组织中均为单等位基因表达,但在体细胞核移植牛的大脑组织中出现了与其他组织亲本来源不同的单等位基因表达。研究结果说明,LINC24065基因在体细胞核移植牛的大脑中出现了印记紊乱,并且剪接体在组织中的表达也发生了改变。以上研究结果说明LINC24065基因与体细胞核移植牛的发育异常有关。
The objective of this study was to analyze the expression profile and imprinting status of a novel long noncoding RNA(LINC24065)located in the DLK1-DIO3 imprinted domain in natural reproduction(NR)and somatic cell nuclear transfer(SCNT)calves,which would provide a theoretical basis for further understanding the role of DLK1-DIO3 imprinted domain in the reprogramming of SCNT.The brain of NR calves was used as experimental material,a novel long intergenic non-coding RNA was identified by RACE and RT-PCR methods and named LINC24065.The expression and imprinting status of LINC24065 in NR and SCNT calves were analyzed by SNP-based direct sequencing of RT-PCR product.Sequence analysis revealed that LINC24065 contained 6 splicing variants which were expressed in 7 detected tissues of NR calves.But in SCNT calves,LINC24065-V3 was not detected and the other 5 isoforms showed tissue-specific expression.Imprinting status analysis indicated that LINC24065 exhibited monoallelic expression in 7 tissues of NR and SCNT calves;but in brain of SCNT calves,the monoallelic expression of LINC24065 had different parent origin from that in other tissues.The aberrant expression patterns and imprinting status of LINC24065 occurred in SCNT calves.The expression of splicing variants also changed in tissues.These results suggest that LINC24065 gene is associated with abnormal development of SCNT cattle.
作者
张萃
陈玮娜
李俊良
谷书凯
徐达
李冬杰
李世杰
ZHANG Cui;CHEN Weina;LI Junliang;GU Shukai;XU Da;LI Dongjie;LI Shijie(College of Life Sciences,Hebei Agricultural University,Baoding 071001,China;College of Traditional Chinese Medicine,Hebei University,Baoding 071002,China;College of Bioscience and Bioengineering,Hebei University of Science and Technology,Shijiazhuang 050018,China)
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2019年第1期44-51,共8页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家自然科学基金(31372312)
河北省自然科学基金(C2016204092)
河北省在读研究生创新能力培养资助项目(CXZZBS2017063)
