牛乳状瘤病毒13型pEGFP-E5-N1真核表达载体的构建及其在HEK293细胞中的表达

doi:10.16431/j.cnki.1671-7236.2015.05.008

Abstract

Abstract: The PCR technology was used to amplify the E5 gene according to the genome sequence of bovine papillomavirus type 13 (BPV-13) Hainan strain with the GenBank accession number KM258443.2.Then it was ligated into pEGFP-N1 vector.The recombinant plasmid pEGFP-E5-N1 was transfected into HEK293 cells with liposomes by transient transfection.The expression of fusion protein was identified by fluorescence microscope, flow cytometry and Western blotting.The results showed that the E5 gene was 135 bp with the 100% homology with the sequence deposited in GenBank.Bright green fluorescence could be seen in HEK293 cells transfected by pEGFP-E5-N1.The expressed fusion protein was about 32 ku.This experiment laid the foundation for further study of the functions of E5 gene in eukaryotic cells.

Key words: bovine papillomavirus type 13 (BPV-13); E5; EGFP; fusion protein; eukaryotic expression

CLC Number:

Q786

PANG Feng, SHI Qiao-yun, ZHU Hua-pei, XU Kai-lian, ZHAO Tian-jing, LI Ya-ying, PENG Dong-mei, LI Guo-hua, ZHOU Hai-long, WANG Feng-yang. Construction of Eukaryotic Expression Vector pEGFP-E5-N1 of Bovine Papillomavirus Type 13 and its Expression in HEK293 Cells[J]. , 2015, 42(5): 1088-1092.

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Construction of Eukaryotic Expression Vector pEGFP-E5-N1 of Bovine Papillomavirus Type 13 and its Expression in HEK293 Cells

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