SLA-Ⅰα链和β链的原核表达与抗血清制备

doi:10.11843/j.issn.0366-6964.2015.07.020

Abstract

Abstract:

The swine major histocompatibility complex class Ⅰ or swine leukocyte antigen class Ⅰ (SLA-Ⅰ)，made of the heavy chain (α chain) and light chain (β chain)，is involved in the process of endogenous antigen presenting.In the present study，we first amplified the genes of the extracellular region in SLA-Ⅰα chain and β chain from porcine alveolar macrophage (PAM) using RT-PCR.The two genes were then cloned into the prokaryotic expression vectors and induced to express in the form of inclusion bodies.After being washed，refolded and purified，the recombinant proteins were confirmed by western blot and mass spectrometer.Then，the rabbit antiserum against the recombinant proteins was prepared with titers of 1：256 000 by an indirect ELISA.Finally，the antiserum was used in Western blot and indirect immunofluorescence assay，showing that the antiserum can specifically react with SLA-Ⅰ α chain and β chain expressed in both PAM and PAM cell line 3D4/21.This study not only provides useful reagents for analyzing the function of SLA-Ⅰ，but also develops the basis for further study on the mechanisms associated with immune responses induced by viral infections.

CLC Number:

S852.4

LIU Ying，DU Ji-ge，GE Xin-na，ZHOU Lei，GUO Xin，YANG Han-chun. Prokaryotic Expression of SLA-Ⅰ α Chain and β Chain and Antiserum Preparation[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(7): 1224-1231.

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Prokaryotic Expression of SLA-Ⅰ α Chain and β Chain and Antiserum Preparation

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