牛NTM基因可变剪接体和印记状态分析  

The Analysis of Splice Variants and Genomic Imprinting Status of NTM Gene in Cattle(Bos taurus)

作　　者：徐达[1] 李俊良 张萃[1] 陈玮娜 李冬杰[3] 李世杰[1] XU Da;LI Jun-Liang;ZHANG Cui;CHEN Wei-Na;LI Dong-Jie;LI Shi-Jie(College of Life Science,Hebei Agricultural University,Baoding 071001,China;Department of Traditional Chinese Medicine,Hebei University,Baoding 071002,China;College of Bioscienee and Bioengineering,Hebei University of Science and Technology,Shijiazhuang 050018,China)

机构地区：[1]河北农业大学生命科学学院,保定071001 [2]河北大学中医系,保定071002 [3]河北科技大学生物科学与工程学院,石家庄050018

出　　处：《农业生物技术学报》2018年第10期1707-1713,共7页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(No.31372312);河北省自然科学基金(No.C2016204092)

摘　　要：NTM (neurotrimin)在神经发育中具有重要作用,在人类(Homo sapiens)相关研究中是印记基因,但是在牛(Bos taurus)中的印记状态尚未揭示。本研究旨在分析牛NTM基因的序列特征和编码蛋白结构,并鉴定NTM基因在成年牛组织和胎盘中的印记状态。本研究首先应用cDNA末端快速扩增技术(rapid amplification of cDNA ends, RACE),得到1个牛NTM基因新的剪接体NTMxd (GenBank No.:MH037574),通过对其编码氨基酸序列进行分析,发现其N端具有完整的跨膜信号。为了分析NTM基因在牛中的印记状态,首先通过PCR产物直接测序法在牛NTM基因上发现1个单核苷酸多态性(single nucleotide polymorphism, SNP)位点,GenBank登录号为rs42185569;然后提取杂合子牛胎盘和组织样本RNA,并进行反转录PCR (reverse transcription-PCR, RT-PCR)。对扩增产物直接测序发现,NTM基因在牛胎盘中和成年牛组织中均为单等位基因表达。本研究结果表明NTM基因在牛中是印记基因,为深入研究NTM基因的功能和相关神经发育疾病提供了参考依据。Neurotrimin（NTM） plays an important role in the nerve development. The NTM is an imprinted gene in human（Homo sapiens）, but the imprinting state in cattle（Bos taurus） has not been revealed. The purpose of present study is to analyze the sequences of cattle NTM gene, the structure of its coding protein,and to identify the imprinting status of NTM gene in adult tissues and placenta in cattle. A new splice NTMxd（GenBank No.： MH037574） of cattle NTM gene was obtained by the technology of rapid amplification of cDNA ends（RACE）, and a more complete transmembrane signal was found in the N-terminal of amino acid sequence encoded by the splice. In order to analyze the imprinting status of NTM gene in cattle, a locus of single nucleotide polymorphism（SNP） was found in cattle NTM gene by direct sequencing of PCR products,the accession number of the SNP locus in NCBI website（www. ncbi. nlm. nih. gov） is rs42185569. The total RNA was extracted from the tissue samples and placentas of heterozygous individuals, and performed expression analysis by reverse transcription-PCR（RT-PCR）. Direct sequencing of the RT-PCR amplified products revealed that NTM gene exhibited monoallelic expression in cattle placenta and adult tissues,indicating that NTM gene is an imprinted gene in cattle. The results of present study provides basic data for further studying of NTM gene function and related neurodevelopmental diseases.

关 键 词：牛 Neurotrimin基因(NTM) 剪接体 印记状态 单核苷酸多态性(SNP) 

分 类 号：S823[农业科学—畜牧学]