猪DNM2基因克

发布时间：2018-05-20 08:46

  本文选题：猪 + DNM基因　； 参考：《中国畜牧兽医》2017年09期

【摘要】：本研究旨在对猪发动蛋白2(dynamin-2,DNM2)基因进行克隆和生物信息学分析,并探讨DNM2基因在猪不同组织中的表达情况。利用RT-PCR结合RACE方法克隆猪DNM2基因cDNA部分序列,与猪表达序列标签进行拼接,获得猪DNM2基因cDNA全长,并对其进行生物信息学分析,同时采用实时荧光定量PCR检测DNM2基因在猪不同组织中的表达情况。结果表明,猪DNM2基因的开放阅读框(open reading fram,ORF)为2 616bp,共编码871个氨基酸。DNM2相对分子质量为98 071.30,等电点(pI)为7.04;无信号肽和跨膜结构域,即该蛋白不属于分泌蛋白;DNM2蛋白的二级结构预测发现,构成α-螺旋、β转角、无规则卷曲、延展链的氨基酸数量分别为361、53、335和122个。多重分析结果显示,猪DNM2基因与牛、人、小鼠、大鼠的序列同源性分别为92.6%、91.8%、88.6%和89.3%;进化树分析表明,DNM2基因在物种间具有较高的保守性,不同物种间DNM2基因序列的差异符合物种间的进化性。实时荧光定量PCR结果显示,DNM2基因在脾脏中表达量较高,在乳腺、腿肌、输卵管、卵巢和子宫中表达量均较低。本研究结果为今后深入研究DNM2基因的生物学功能奠定了基础。
[Abstract]:The purpose of this study was to clone and bioinformatics analysis of porcine motor protein 2dynamin-2 (DNM2) gene, and to investigate the expression of DNM2 gene in different tissues of pigs. The partial cDNA sequence of porcine DNM2 gene was cloned by RT-PCR combined with RACE method and spliced with porcine expression sequence tag to obtain the full length of porcine DNM2 gene cDNA and its bioinformatics analysis. Real-time quantitative PCR was used to detect the expression of DNM2 gene in different pig tissues. The results showed that the open reading frame of porcine DNM2 gene was 2 616 BP, encoding 871 amino acids. DNM2 had a molecular weight of 98 071.30 and an isoelectric point (Pi) of 7. 04, no signal peptide and transmembrane domain. That is to say, the protein does not belong to the secondary structure of secretory protein DNM2. It is found that the protein forms 伪 -helix, 尾 rotation angle, irregular crimp, and the number of amino acids extending the chain is 361n 53335 and 122, respectively. The results of multiple analysis showed that the sequence homology of porcine DNM2 gene with cattle, human, mouse and rat was 91.6% and 89.3%, respectively, and the phylogenetic tree analysis showed that DNM2 gene was highly conserved among species. The difference of DNM2 gene sequence between different species is consistent with the evolution of different species. The results of real-time fluorescence quantitative PCR showed that the expression of DNM2 gene in spleen was higher than that in breast, leg muscle, fallopian tube, ovary and uterus. The results laid a foundation for the further study of the biological function of DNM2 gene.
【作者单位】： 浙江农林大学动物科技学院;
【基金】：国家自然科学基金(31501921) 浙江省自然科学基金(LQ15C170001) 浙江省农业(畜禽)新品种选育重大科技专项(2016C02054-3)
【分类号】：Q78;S828
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本文编号：1913948