猪圆环病毒2型CAP蛋白在flashBAC杆状病毒表达系统中的表达与鉴定

doi:10.16431/j.cnki.1671-7236.2016.07.033

Abstract

Abstract:

The study was aimed to express CAP protein of porcine circovirus type 2 (PCV2) by eukaryotic expression system.The PCV2 TZ0601 strain was the template,the CAP protein with or without the signal peptide of PCV2 coding sequence were cloned into pOET3 vector.The constructed plasmid were confirmed by restriction enzyme digestion and DNA sequencing,then sf9 insect cells were transfected with recombinant plasmid pOET3-CAP and pOET3-CAP-X.The test was designed to express the CAP protein with or without the signal peptide of PCV2 by flashBAC baculovirus expression system.Expression of PCV2 gene were confirmed by indirect immunofluorescent assay (IFA),SDS-PAGE and Western blotting.The results showed that the eukaryotic expression plasmids of pOET3-CAP and pOET3-CAP-X were constructed successfully and the gene was highly expressed in sf9 insect cells.After expression,we could see our target band about 25 to 35 ku with SDS-PAGE and Western blotting.The immuno-reactivity of the protein was confirmed by antisera against PCV2.This would lay a foundation for a further study of PCV2 subunit vaccine and diagnostic antigen kit.

Key words: porcine circovirus type 2; flashBAC baculovirus expression system; CAP protein

CLC Number:

S852.65⁺9.2

GUO Hui-juan, FAN Cui, ZHANG Ying, LV Mao-jie, LIANG Wu, YANG Bao-shou. Expression and Identification of Porcine Circovirus type 2 Capsid Protein Using flashBAC Baculovirus Expression System[J]. , 2016, 43(7): 1876-1883.

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Expression and Identification of Porcine Circovirus type 2 Capsid Protein Using flashBAC Baculovirus Expression System

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