东北白鹅CD4基因的克隆及其胞外区的表达与抗血清的制备

doi:10.11843/j.issn.0366-6964.2014.04.019

Abstract

Abstract:

The aim of this study was to prepare antiserum against the goose CD4 protein.Specific primers for goose CD4 gene were designed according to public Mallard duck reference sequence (AF378701) in GenBank.Then we obtained Northeast White Goose CD4 gene by RT-PCR successfully.According to goose CD4 ORF sequence，specific primers were further designed and goose CD4 extracellular region gene was cloned.We produced recombinant goose CD4 extracellular region protein in prokaryotic expression system (pET-32a（+）/Rosetta(DE3)pLysS).The recombinant protein was induced by IPTG and was purified by Ni-NTA column affinity chromatography.As immunogen，the purified recombinant protein prepared rabbit anti-goose CD4 extracellular region serum.I-ELISA，Western blot and Flow cytometric analysis showed that antiserum could identify recombinant goose CD4 extracellular region protein and peripheral blood T lymphocytes respectively.Indirect immunofluorescence test confirmed the purified antiserum could identify goose CD4 extracellular protein by eukaryotic expression system specifically.The above results show that antiserum can be used as detection reagent to detect goose CD4⁺ T lymphocytes.

CLC Number:

S852.4

ZHANG Xue-lian，WEI Shuang-shi，SHAO Jian-wei，MU Xiao-yu，GAO Ming-chun，ZHANG Wen-long，MA Bo，WANG Jun-wei. Molecular Cloning and Expression of Extracellular Region of Northeast White Goose CD4 Gene and Preparation of Its Antiserum[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(4): 639-646.

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Molecular Cloning and Expression of Extracellular Region of Northeast White Goose CD4 Gene and Preparation of Its Antiserum

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