本文選題：豬 + DNM基因　； 參考：《中國畜牧獸醫(yī)》2017年09期

【摘要】：本研究旨在對(duì)豬發(fā)動(dòng)蛋白2(dynamin-2,DNM2)基因進(jìn)行克隆和生物信息學(xué)分析,并探討DNM2基因在豬不同組織中的表達(dá)情況。利用RT-PCR結(jié)合RACE方法克隆豬DNM2基因cDNA部分序列,與豬表達(dá)序列標(biāo)簽進(jìn)行拼接,獲得豬DNM2基因cDNA全長,并對(duì)其進(jìn)行生物信息學(xué)分析,同時(shí)采用實(shí)時(shí)熒光定量PCR檢測(cè)DNM2基因在豬不同組織中的表達(dá)情況。結(jié)果表明,豬DNM2基因的開放閱讀框(open reading fram,ORF)為2 616bp,共編碼871個(gè)氨基酸。DNM2相對(duì)分子質(zhì)量為98 071.30,等電點(diǎn)(pI)為7.04;無信號(hào)肽和跨膜結(jié)構(gòu)域,即該蛋白不屬于分泌蛋白;DNM2蛋白的二級(jí)結(jié)構(gòu)預(yù)測(cè)發(fā)現(xiàn),構(gòu)成α-螺旋、β轉(zhuǎn)角、無規(guī)則卷曲、延展鏈的氨基酸數(shù)量分別為361、53、335和122個(gè)。多重分析結(jié)果顯示,豬DNM2基因與牛、人、小鼠、大鼠的序列同源性分別為92.6%、91.8%、88.6%和89.3%;進(jìn)化樹分析表明,DNM2基因在物種間具有較高的保守性,不同物種間DNM2基因序列的差異符合物種間的進(jìn)化性。實(shí)時(shí)熒光定量PCR結(jié)果顯示,DNM2基因在脾臟中表達(dá)量較高,在乳腺、腿肌、輸卵管、卵巢和子宮中表達(dá)量均較低。本研究結(jié)果為今后深入研究DNM2基因的生物學(xué)功能奠定了基礎(chǔ)。
[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 偽 -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.
【作者單位】： 浙江農(nóng)林大學(xué)動(dòng)物科技學(xué)院;
【基金】：國家自然科學(xué)基金(31501921) 浙江省自然科學(xué)基金(LQ15C170001) 浙江省農(nóng)業(yè)(畜禽)新品種選育重大科技專項(xiàng)(2016C02054-3)
【分類號(hào)】：Q78;S828
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本文編號(hào)：1913948