LINC24065在体细胞核移植牛中的异常表达

doi:10.11843/j.issn.0366-6964.2019.01.006

畜牧兽医学报 ›› 2019, Vol. 50 ›› Issue (1): 44-51.doi: 10.11843/j.issn.0366-6964.2019.01.006

LINC24065在体细胞核移植牛中的异常表达

张萃¹, 陈玮娜^1,2, 李俊良¹, 谷书凯¹, 徐达¹, 李冬杰³, 李世杰^1*

1. 河北农业大学生命科学学院, 保定 071001;
2. 河北大学中医学院, 保定 071002;
3. 河北科技大学生物科学与工程学院, 石家庄 050018

收稿日期:2018-05-14 出版日期:2019-01-23 发布日期:2019-01-23
通讯作者: 李世杰,主要从事动物分子遗传研究,E-mail:lishijie20005@163.com
作者简介:张萃(1976-),女,河北辛集人,博士,主要从事动物分子遗传研究,E-mail:zhangcui@hebau.edu.cn;陈玮娜(1980-),女,河北保定人,博士,主要从事动物分子遗传研究,E-mail:chenweina429@163.com
基金资助:
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）

Abnormal Expression of LINC24065 in Somatic Cell Nuclear Transfer Cattle

ZHANG Cui¹, CHEN Weina^1,2, LI Junliang¹, GU Shukai¹, XU Da¹, LI Dongjie³, LI Shijie^1*

1. College of Life Sciences, Hebei Agricultural University, Baoding 071001, China;
2. College of Traditional Chinese Medicine, Hebei University, Baoding 071002, China;
3. College of Bioscience and Bioengineering, Hebei University of Science and Technology, Shijiazhuang 050018, China

Received:2018-05-14 Online:2019-01-23 Published:2019-01-23

摘要/Abstract

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。

Abstract:

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.

中图分类号:

S823
S813.3

张萃, 陈玮娜, 李俊良, 谷书凯, 徐达, 李冬杰, 李世杰. LINC24065在体细胞核移植牛中的异常表达[J]. 畜牧兽医学报, 2019, 50(1): 44-51.

ZHANG Cui, CHEN Weina, LI Junliang, GU Shukai, XU Da, LI Dongjie, LI Shijie. Abnormal Expression of LINC24065 in Somatic Cell Nuclear Transfer Cattle[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(1): 44-51.

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LINC24065在体细胞核移植牛中的异常表达

Abnormal Expression of LINC24065 in Somatic Cell Nuclear Transfer Cattle

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