Genetic mutations potentially cause two novel NCF1 splice variants up-regulated in the mammary gland, blood and neutrophil of cows infected by Escherichia coli.
Genetic mutations potentially cause two novel NCF1 splice variants up-regulated in the mammary gland, blood and neutrophil of cows infected by Escherichia coli.
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基因突变可能导致受大肠杆菌感染的奶牛的乳腺、血液和中性粒细胞中两种新的 NCF1 剪接变体上调。
DOI：
10.1016/j.micres.2015.03.005
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发表时间：
2015-05
影响因子：
6.7
通讯作者：
Huang Jinming
中科院分区：
文献类型：
--
作者：
Zhang Zijing;Wang Xiuge;Li Rongling;Ju Zhihua;Qi Chao;Zhang Yan;Guo Fang;Luo Guojing;Li Qiuling;Wang Changfa;Zhong Jifeng;Xu Yinxue;Huang Jinming
中性粒细胞胞浆因子1(NCF1)在宿主抵抗病原体的过程中起着至关重要的作用。在这项研究中，我们研究了牛NCF1基因的潜在选择性剪接模式、表达和剪接相关的单核苷酸多态(SNPs)，以增加对其对由大肠杆菌感染引起的牛乳房炎的潜在作用的认识。利用RT-PCR和克隆测序的方法，我们发现了两个新的剪接变异体，分别设计为NCF1-TV1(保留内含子6)和NCF1-TV2(保留内含子8的部分)，编码两个推测的截短蛋白(239AA和283AA)。实时定量RT-PCR检测发现，与健康奶牛相比，乳房炎感染奶牛的乳腺组织、血液和中性粒细胞中两种剪接变异体的表达显著上调。基因组测序分析在NCF1基因内含子6和内含子8末端发现了4个新的SNPs g.10112G>；A、g.10766T>；C、g.12085G>；A和g.12430T>；C。ESE基序预测SNP(g.10766T>；C)可能影响与剪接相关因子的结合，从而导致异常剪接变异体NCF1-TV1的产生，这是功能性SNP与奶牛乳体细胞评分增加相关的潜在原因之一。我们的结果将有助于更好地理解NCF1基因在抗病原体感染过程中的功能，以及剪接相关SNP对奶牛异常剪接变体产生的影响和仔细的功能鉴定。
Neutrophil cytosolic factor 1(NCF1) plays a crucial role in host defense against microbial pathogens. In this study, we investigated the potential alternative splicing patterns, expression and splice-relevant single nucleotide polymorphisms (SNPs) of the bovineNCF1gene to increase insights into its potential role against bovine mastitis caused byEscherichia coliinfection. Using RT-PCR and clone sequencing methods, we found two novel splice variants designed asNCF1-TV1(retained intron 6) andNCF1-TV2(retained part of intron 8), respectively, encoding two putative truncated proteins (239AA and 283AA). Two splice variants were drastically up-regulated in the mastitis-infected cows’ mammary tissues, blood and neutrophils compared with these of healthy cows using real-time RT-PCR. Genomic sequencing analysis identified four novel SNPs g.10112 G>A, g.10766 T>C, SNPs g.12085 G>A and g.12430 T>C at the ends of intron 6 and intron 8 ofNCF1. ESE motif predicted that the SNP (g.10766 T>C) might affect the binding with splicing-related factors and subsequently caused the production of aberrant splice variantNCF1-TV1, which is one of the potential reasons that the functional SNP was associated with increased milk somatic cell score in cow. Our results would help in better understanding theNCF1gene function in the process against pathogen infection, and the effect of splicing-related SNP on the production of aberrant splice variant and careful functional characterization in dairy cattle.
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影响因子：
44.1
作者：
Shi, YY;He, L
通讯作者：
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作者：
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Sharp PA
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作者：
Ju, Zhihua;Wang, Changfa;Zhong, Jifeng
通讯作者：
Zhong, Jifeng