猪JHDM2A基因的克隆及表达分析

发布时间：2018-03-05 16:35

  本文选题：猪　切入点：JHDMA基因　出处：《中国畜牧兽医》2017年06期 　论文类型：期刊论文

【摘要】：试验旨在克隆猪JHDM2A基因,并研究其在猪卵巢组织中的表达情况。首先克隆猪JHDM2A基因,并构建pLVX-IRES-ZsGreen1-JHDM2A真核表达载体,同时对JHDM2A基因在猪卵泡发育过程中的表达情况进行分析。结果显示,克隆得到的猪JHDM2A基因编码区长度为3 945bp,编码1 315个氨基酸。通过多重氨基酸序列比对发现,猪JHDM2A基因与黄牛、水牛、绵羊和人相应氨基酸序列的同源性分别为93.5%、94.7%、94.7%和93.8%。蛋白质分子系统进化树分析结果表明,JHDM2A基因在物种进化过程中高度保守。通过脂质体转染法将构建的pLVX-IRES-ZsGreen1-JHDM2A真核表达载体导入HEK293T细胞,可观察到清晰的绿色荧光蛋白表达。免疫组化结果显示,JHDM2A蛋白在不同发育阶段猪卵泡中均有表达。本试验通过克隆获得猪JHDM2A基因序列,JHDM2A蛋白在猪卵巢中高度表达,表明其功能可能与猪卵泡发育密切相关。
[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.
【作者单位】： 广西大学亚热带农业生物资源保护与利用国家重点实验室;
【基金】：国家自然科学基金(31560632) 广西自然科学基金项目(2014GXNSFAA118099、2016GXNSFDA380030)
【分类号】：Q78;S828
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本文编号：1571050