摘要
RNF4基因调节精细胞分化和增殖过程,文章以Gen Bank下载的猪及近缘物种RNF4 mRNA序列为参考序列,设计特异引物扩增版纳微型猪近交系(BMI)RNF4基因。应用qPCR分析15个重要组织mRNA表达谱,并对蛋白质序列作功能生物信息学分析,构建多物种系统进化树。研究获得BMI RNF4 573 bp编码区序列(Gen Bank登录号:KU705638,对应氨基酸登录号:AOC89056),编码190个氨基酸,蛋白质分子质量(Mw)为21.43 ku,等电点(pI)为6.95。多组织荧光定量表达分析表明RNF4基因在尿道球腺、睾丸、肝、肺中高水平表达;在其他11个组织中呈中低水平表达。功能生物信息学分析表明RNF4蛋白质存在1个保守结构域,无跨膜结构,无信号肽序列;N末端疏水,C末端疏水;有4类功能活性位点,位于细胞核概率为94.1%。系统进化分析表明,BMI与狗亲缘关系最近。结果为研究RNF4基因在猪精细胞分化和增殖方面作用奠定基础。
RNF4 gene plays a regulater of sperm cell differentiation and proliferation process. The RNF4 mRNA sequence of pig and related species from GenBank was used as reference sequences, We designed specific primers and amplified Banna mini-pig inbred line （BMI） RNF4 gene. qPCR was applied to analysis the mRNA expression profiles of 15 important tissues. Protein sequence was used to carry out functional bioinformatics analysis and construct phylogenetic tree. A coding sequence of 573 bp （GenBank accession number： KU705638, the corresponding amino acid sequence accession number： AOC89056） of BMI RNF4 was obtained, which encodes a protein of 190 amino acids, protein molecular weight 21.43 ku, and isoelectric point 6.95. Fluorescence quantitative expression indicated that RNF4 gene expressed highly in the antiprostate, testis, liver, lung, and low expression in the other 11 tissues. Functional bioinformaUcs analysis indicated that RNF4 protein contained one conserved domains, no transmembrane region, no signal peptide sequences; its N-terminal and C-terminal were hydrophobic; it has four kinds functional active sites and a percentage 94.1 to located in cell nucleus. Phylogenetic analysis demonstrated that BMI had the closest relationship with dog. The results of the study will lay a foundation for further study of the gene about its functions in pig sperm cell differentiation and proliferation.
作者
张霞
曾日彬
霍金龙
王配
陈园园
王淑燕
ZHANG Xia ZENG Ribin HUO Jinlong WANG Pei CHEN Yuanyuan WANG Shuyan(Key Laboratory of Banna Mini-pig Inbred Line of Yunnan Province, Yunnan Agricultural University, Kunming 650201, China School of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, China)
出处
《东北农业大学学报》
CAS
CSCD
北大核心
2017年第3期24-30,共7页
Journal of Northeast Agricultural University
基金
国家自然科学基金项目(31660637
31460580
31660650)
