本文選題：豬　切入點(diǎn)：JHDMA基因　出處：《中國(guó)畜牧獸醫(yī)》2017年06期 　論文類(lèi)型：期刊論文

【摘要】：試驗(yàn)旨在克隆豬JHDM2A基因,并研究其在豬卵巢組織中的表達(dá)情況。首先克隆豬JHDM2A基因,并構(gòu)建pLVX-IRES-ZsGreen1-JHDM2A真核表達(dá)載體,同時(shí)對(duì)JHDM2A基因在豬卵泡發(fā)育過(guò)程中的表達(dá)情況進(jìn)行分析。結(jié)果顯示,克隆得到的豬JHDM2A基因編碼區(qū)長(zhǎng)度為3 945bp,編碼1 315個(gè)氨基酸。通過(guò)多重氨基酸序列比對(duì)發(fā)現(xiàn),豬JHDM2A基因與黃牛、水牛、綿羊和人相應(yīng)氨基酸序列的同源性分別為93.5%、94.7%、94.7%和93.8%。蛋白質(zhì)分子系統(tǒng)進(jìn)化樹(shù)分析結(jié)果表明,JHDM2A基因在物種進(jìn)化過(guò)程中高度保守。通過(guò)脂質(zhì)體轉(zhuǎn)染法將構(gòu)建的pLVX-IRES-ZsGreen1-JHDM2A真核表達(dá)載體導(dǎo)入HEK293T細(xì)胞,可觀察到清晰的綠色熒光蛋白表達(dá)。免疫組化結(jié)果顯示,JHDM2A蛋白在不同發(fā)育階段豬卵泡中均有表達(dá)。本試驗(yàn)通過(guò)克隆獲得豬JHDM2A基因序列,JHDM2A蛋白在豬卵巢中高度表達(dá),表明其功能可能與豬卵泡發(fā)育密切相關(guān)。
[Abstract]:The aim of this study was to clone porcine JHDM2A gene and study its expression in porcine ovary. Firstly, porcine JHDM2A gene was cloned and eukaryotic expression vector of pLVX-IRES-ZsGreen1-JHDM2A was constructed. At the same time, the expression of JHDM2A gene in the development of pig follicle was analyzed. The results showed that the encoding region of the cloned porcine JHDM2A gene was 3 945 BP, encoding 1 315 amino acids, and was found by multiple amino acid sequence alignment. Pig JHDM2A gene and yellow cattle, water buffalo, The homology of corresponding amino acid sequences between sheep and human was 94.5% and 93.8%, respectively. The results of phylogenetic tree analysis showed that the JHDM2A gene was highly conserved in the evolution of species. The constructed pLVX-IRES-ZsGreen1-JHDM2A was expressed in eukaryotic by liposome transfection. The vector was introduced into HEK293T cells. A clear expression of green fluorescent protein was observed. Immunohistochemical results showed that JHDM2A protein was expressed in pig follicles at different stages of development. In this study, the pig JHDM2A gene sequence, JHDM2A protein, was highly expressed in porcine ovaries. It suggests that its function may be closely related to the development of pig follicles.
【作者單位】： 廣西大學(xué)亞熱帶農(nóng)業(yè)生物資源保護(hù)與利用國(guó)家重點(diǎn)實(shí)驗(yàn)室;
【基金】：國(guó)家自然科學(xué)基金(31560632) 廣西自然科學(xué)基金項(xiàng)目(2014GXNSFAA118099、2016GXNSFDA380030)
【分類(lèi)號(hào)】：Q78;S828
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本文編號(hào)：1571050