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首页> 外文会议>International Conference on Bioinformatics and Biomedical Technology >Construction of SMYD3 Liver-specific Expression Transgene for the Establishment of Transgenic Mice
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Construction of SMYD3 Liver-specific Expression Transgene for the Establishment of Transgenic Mice

机译：SMYD3肝脏特异性表达转基因的建立，用于建立转基因小鼠

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摘要

SET and MYND domain-containing protein 3 (SMYD3) is a novel histone methyltransferase involved in human carcinogenesis. To further establish the transgenic mice of SMYD3 and provide a useful platform for the study of SMYD3 and the screening of novel antitumor drugs, a transgene for liver-specific simultaneous expression of SMYD3 and EGFP was constructed. The human SMYD3 gene was subcloned into pIRES2-EGFP eukaryotic expressing vector. Subsequently, mice albumin promoter (pALB) (-821～+59) was obtained from the mice genome DNA by PCR and then inserted into the 5' flanking region of SMYD3. The final recombinant plasmid (named as pIRAS) was digested by AflII StuI and XhoI, and the DNA fragment consisting of albumin promoter (pALB), SMYD3, Internal Ribosome Entry Site (IRES), Enhanced Green Fluorescent Protein (EGFP) and SV40 polyA signal sequence (named as ASIES) was released. The linearized, purified ASIES transgene was microinjected into C57BL/6 mouse zygotes. After cultured for 2 days in vitro, most of the injected zygotes showed normal division, and the following works would establish the transgenic mice.
机译：含有和MyND域的蛋白质3（Smyd3）是一种参与人致癌作用的新型组蛋白甲基转移酶。为了进一步建立Smyd3的转基因小鼠并为SMYD3的研究提供有用的平台，并制成了新型抗肿瘤药物的筛选，构建了用于肝脏特异性Smyd3和EGFP的转基因。人体Smyd3基因亚克隆到Pires2-EGFP真核表达载体中。随后，通过PCR从小鼠基因组DNA获得小鼠白蛋白启动子（PALB）（-821〜+ 59），然后插入Smyd3的5'侧翼区域中。通过AFLII STUI和XHOI消化最终重组质粒（命名为Piras），以及由白蛋白启动子（PALB），SMYD3，内核糖体进入部位（IRES），增强的绿色荧光蛋白（EGFP）和SV40多元信号组成的DNA片段释放序列（命名为ASIES）。线性化纯化的ASIES转基因微量注射到C57BL / 6小鼠Zygotes中。在体外培养2天后，大多数注射的Zygotes显示正常分裂，下列作品将建立转基因小鼠。

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