本文選題：牦牛 + 內(nèi)皮素��； 參考：《獸類學(xué)報(bào)》2017年03期

【摘要】：Endothelin-1主要通過促進(jìn)血管平滑肌細(xì)胞增殖引起血管結(jié)構(gòu)和功能的改變,從而刺激血管收縮,Endothelin-1在動(dòng)物血管生成和低氧適應(yīng)等關(guān)鍵發(fā)育和生理過程中發(fā)揮重要的作用。為了進(jìn)一步探討高原動(dòng)物腦Endothelin-1對(duì)其極端低氧適應(yīng)調(diào)控的分子機(jī)理,本研究對(duì)牦牛(Bos grunniens)腦Endothelin-1基因CDS全長序列進(jìn)行了克隆及其真核表達(dá)載體的構(gòu)建。結(jié)果表明,牦牛Endothelin-1基因的CDS全長序列含有一個(gè)609 bp的開放閱讀框,編碼202個(gè)氨基酸,該蛋白分子量為22.98 k Da,理論等電點(diǎn)(p I)為9.63;成功構(gòu)建出帶有CMV啟動(dòng)子,綠色熒光蛋白標(biāo)記的牦牛Endothelin-1真核表達(dá)質(zhì)粒p EGFP-C1-Endothelin-1;牦牛Endothelin-1基因CDS全長序列編碼的氨基酸序列與黃牛(Bos taurus)、藏羚羊(Pantholop shodgsoni)、綿羊(Ovis aries)、野豬(Sus scrofa)、小鼠(Mus musculus)、人(Homo sapiens)的同源性分別為100%、95%、93%、81%、77%、70%;且該氨基酸序列的系統(tǒng)進(jìn)化情況與其親緣關(guān)系遠(yuǎn)近一致。因此,我們認(rèn)為高原動(dòng)物牦牛Endothelin-1的結(jié)構(gòu)和功能在進(jìn)化上均具有高度保守性,其在低氧適應(yīng)中的作用可能是通過表達(dá)差異或轉(zhuǎn)錄后修飾實(shí)現(xiàn)的,且本研究構(gòu)建的真核表達(dá)質(zhì)粒為進(jìn)一步探討該基因在青藏高原動(dòng)物腦對(duì)極端低氧生境適應(yīng)中的生物學(xué)功能及其調(diào)控機(jī)理提供了支持。
[Abstract]:Endothelin-1 plays an important role in the development and physiological process of angiogenesis and hypoxic adaptation by promoting the proliferation of vascular smooth muscle cells and causing the changes of vascular structure and function, thus stimulating the vasoconstriction of Endothelin-1. In order to further investigate the molecular mechanism of brain Endothelin-1 regulating extremely hypoxic adaptation in plateau animals, the full-length CDS sequence of the brain Endothelin-1 gene of Bos grunniens was cloned and its eukaryotic expression vector was constructed. The results showed that the full-length CDS sequence of yak Endothelin-1 gene contained a 609bp open reading frame encoding 202 amino acids, the molecular weight of the protein was 22.98kDaand the theoretical isoelectric point (Ip) was 9.63. the promoter with CMV was successfully constructed. The eukaryotic expression plasmid pEGFP-C1-Endothelin-1of yak Endothelin-1 labeled with green fluorescent protein; the amino acid sequence encoded by the full-length CDS sequence of yak Endothelin-1 gene is homologous to Bos taurusus, Pantholop shodgsonike, Ovis ariesus, Sus scrofaa, Mus musculus, Homo sapiensus, respectively. The phylogeny of the amino acid sequence is consistent with its phylogenetic relationship. Therefore, we believe that the structure and function of Endothelin-1 in plateau animals are highly conserved in evolution, and its role in hypoxic adaptation may be achieved by expression difference or post-transcriptional modification. The eukaryotic expression plasmid was constructed in this study to further study the biological function and regulation mechanism of the gene in the animal brain of Qinghai-Xizang Plateau to adapt to extremely hypoxic habitat.
【作者單位】： 蘭州大學(xué)生命科學(xué)學(xué)院動(dòng)物學(xué)與發(fā)育生物學(xué)研究所;西北師范大學(xué)生命科學(xué)學(xué)院;
【基金】：國家自然科學(xué)基金項(xiàng)目(31000190,051000006,31060141)
【分類號(hào)】：Q953

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