Construction and Identification of Recombinant Eukaryotic Plasmid of Sika Deer Thymosin Beta10

doi:10.13560/j.cnki.biotech.bull.1985.2016-1000

Abstract

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.

Key words: sika deer, thymosin beta10, plasmid construction, transfection, eukaryotic expression

WANG Jia-wen, GAO Yun-peng, SUN Tian-xia, LIU Mei-chen, ZHAO Yu, WANG Si-ming. Construction and Identification of Recombinant Eukaryotic Plasmid of Sika Deer Thymosin Beta10[J]. Biotechnology Bulletin, 2017, 33(6): 149-154.

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