期刊
MICROBIOLOGICAL RESEARCH
卷 174, 期 -, 页码 24-32出版社
ELSEVIER GMBH
DOI: 10.1016/j.micres.2015.03.005
关键词
NCF1 gene; Aberrant splicing; Functional SNP; Mastitis; Cattle
类别
资金
- National Natural Science Foundation of China [31371255, 31271328, 31000543]
- Support Program of the Ministry of Science and Technology the Major Project of National Transgene in China
- Program of National Cow Industrial Technology System [CARS-37]
Neutrophil cytosolic factor I (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 bovine NCF1 gene to increase insights into its potential role against bovine mastitis caused by Escherichia coli infection. Using RT-PCR and clone sequencing methods, we found two novel splice variants designed as NCF1-TV1 (retained intron 6) and NCF1-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 of NCF1. 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 variant NCF1-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 the NCF1 gene 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. (C) 2015 Elsevier GmbH. All rights reserved.
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