梅花鹿胸腺素beta10基因真核表达载体的构建及鉴定

发布时间：2018-04-29 05:18

  本文选题：梅花鹿 + 胸腺素beta　； 参考：《生物技术通报》2017年06期

【摘要】：转录组测序发现Tβ10在鹿茸中高表达,为了研究Tβ10与鹿茸生长发育间的关系,构建Tβ10真核表达载体。使用Trizol法提取梅花鹿鹿茸总RNA,PCR技术特异性扩增梅花鹿Tβ10基因,利用酶切位点将目的片段插入真核表达载体VR1012构建表达质粒,通过Fugene~?6将质粒瞬时转染到293T细胞中,使用Western blotting和免疫荧光方法检测目的基因的表达。结果发现克隆得到的梅花鹿Tβ10基因长度为129bp,编码42个氨基酸,成功构建真核表达载体VR1012-Tβ10-HA,转染后使用Western blotting方法检测到梅花鹿Tβ10在293T细胞中表达,免疫荧光方法证明梅花鹿Tβ10主要定位在细胞浆中。
[Abstract]:In order to study the relationship between T 尾 10 and the growth and development of velvet antler, the eukaryotic expression vector of T 尾 10 was constructed. The T 尾 10 gene of sika deer was amplified by Trizol. The target fragment was inserted into eukaryotic expression vector VR1012 to construct the expression plasmid. The plasmid was transiently transfected into 293T cells by Fugene~?6. Western blotting and immunofluorescence were used to detect the expression of the target gene. The results showed that the length of T 尾 10 gene of sika deer was 129 BP, encoding 42 amino acids, and the eukaryotic expression vector VR1012-T 尾 10 HA was successfully constructed. After transfection, the expression of T 尾 10 in 293T cells was detected by Western blotting. Immunofluorescence assay showed that T 尾 10 was mainly localized in the cytoplasm of sika deer.
【作者单位】： 长春中医药大学中医药与生物工程研究开发中心;
【基金】：吉林省科技发展计划(20140622003JC)
【分类号】：Q78;S825
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本文编号：1818567