生物技术通报2017，Vol.33Issue(6)：149-154,6.DOI:10.13560/j.cnki.biotech.bull.1985.2016-1000
梅花鹿胸腺素beta10基因真核表达载体的构建及鉴定
Construction and Identification of Recombinant Eukaryotic Plasmid of Sika Deer Thymosin Beta10
摘要
Abstract
Tβ10 was discovered to be highly expressed in sika antler in transcriptome sequencing. In order to study the relationship between Tβ10 and the growth and development of antler,the Tβ10 eukaryotic expression vector was constructed. Extracting the total RNA from sika deer antler by Trizol regent,the coding region of Tβ10 gene was amplified by RT-PCR,and then the gene fragment was inserted into the recombinant eukaryotic expression vector VR1012. The eukaryotic expression plasmids were transfected into 293T cells by Fugene 6 agent. The ORF of Tβ10 cDNA was 129 bp,encoding for a protein of 42 amino acids. The eukaryotic expression vector VR1012- Tβ10-HA was constructed successfully. The expression of sika deer Tβ10 was detected in 293T cells by Western blotting. Immunofluorescence demonstrated that the expressed fusion protein of the Tβ10 was located in the cytoplasm in 293T cells.关键词
梅花鹿/胸腺素beta10/质粒构建/转染/真核表达Key words
sika deer/thymosin beta10/plasmid construction/transfection/eukaryotic expression引用本文复制引用
王佳雯,高云鹏,孙天霞,刘美辰,赵雨,王思明..梅花鹿胸腺素beta10基因真核表达载体的构建及鉴定[J].生物技术通报,2017,33(6):149-154,6.基金项目
吉林省科技发展计划(20140622003JC) （20140622003JC）
