新型鸭呼肠孤病毒σB蛋白的原核表达及其多克隆抗体制备

doi:10.16431/j.cnki.1671-7236.2016.12.010

Abstract

Abstract:

To prepare polyclonal antibodies against σB protein of new-type duck reovirus (NDRV) XX strain, the encoding sequence of σB gene of NDRV XX strain was amplified by RT-PCR and successfully inserted to expression plasmid pET-32a(+), and transformed in Escherichia coli BL21(DE3). The His-σB recombinant protein was achieved with IPTG induction. SDS-PAGE result showed that the molecular weight of the expression on fusion protein was about 55 ku, was major insoluble fractions. IPTG induced time and concentration were 3 h and 0.25 mmol/L, respectively. The soluble σB recombinant protein was highly purified which was purified using Ni²⁺ affinity chromatography and verified by Western blotting and protein mass spectrometry. Then the polyclonal antibodies could be obtained from the rabbits which had immunized by the purified σB recombinant protein with the reasonable procedure. The Western blotting result showed that they had the specific reaction. The results built a foundation of the further study of the NDRV σB protein function and the research of genetic engineering vaccine.

Key words: new-type duck reovirus; σB protein; prokaryotic expression; polyclonal antibody

CLC Number:

S852.65⁺9.4

MEI Min-min, LIANG Guo-zhi, HUANG Wen-jing, LI Xiao-wen, HUANG Shu-jian. Prokaryotic Expression and Polyclonal Antibody Preparation of σB Protein of New-type Duck Reovirus[J]. , 2016, 43(12): 3149-3155.

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Prokaryotic Expression and Polyclonal Antibody Preparation of σB Protein of New-type Duck Reovirus

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