真核表达载体pIRES2-AcGFP1-PLIN2的构建及其在3T3-L1前脂肪细胞中的表达

doi:10.19640/j.cnki.jtau.2018.02.014

Journal of Tianjin Agricultural University ›› 2018, Vol. 25 ›› Issue (2): 55-59.doi: 10.19640/j.cnki.jtau.2018.02.014

• Researches and Scientific Notes • Previous Articles Next Articles

Construction of eukaryotic expression plasmid pIRES2-AcGFP1- PLIN2 and its expression in 3T3-L1 preadipocytes

ZHAO Xiao-qi, HUANG Ying, YANG Ming-hua, PAN Hong-bin, ZHAO Su-mei^{Corresponding Author}

Key Lab of Agricultural Animal Nutrition and Feed Science, Yunnan Agricultural University, Kunming 650201, China

Received:2018-03-26 Online:2018-06-20 Published:2019-11-12

Abstract

Abstract: The eukaryotic expression plasmid pIRES2-AcGFP1-PLIN2 was constructed and its transfection status in 3T3-L1 preadipocytes was observed. The PLIN2 gene was cloned into pMD18-T vector and digested by EcoR I/Sal I. The digested product was digested and then ligated into pIRES2-AcGFP1 vector and transformed into DH5α competent cells, and pIRES2-AcGFP1-PLIN2 positive recombinant plasmid was extracted and transfected into 3T3-L1 preadipocytes. The Transfection condition was observed by microscope. The PLIN2 gene was successfully cloned. At the same time, the eukaryotic expression plasmid pIRES2-AcGFP1-PLIN2 was successfully constructed and expressed in 3T3-L1 preadipocytes. The green fluorescence was observed under the microscope, which laid the foundation for further studying the regulation mechanism of PLIN2 gene on adipocytes.

Key words: Wujin pig, PLIN2 gene, eukaryotic expression plasmid

CLC Number:

S811.3

ZHAO Xiao-qi, HUANG Ying, YANG Ming-hua, PAN Hong-bin, ZHAO Su-mei. Construction of eukaryotic expression plasmid pIRES2-AcGFP1- PLIN2 and its expression in 3T3-L1 preadipocytes[J]. Journal of Tianjin Agricultural University, 2018, 25(2): 55-59.

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Construction of eukaryotic expression plasmid pIRES2-AcGFP1- PLIN2 and its expression in 3T3-L1 preadipocytes

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