畜牧与兽医2017，Vol.49Issue(9)：67-70,4.
禽网状内皮组织增生症病毒CA蛋白的表达纯化及多克隆抗体制备
Expression and purification of reticuloendotheliosis virus CA protein and preparation of polyclonal antibody
摘要
Abstract
To express the CA protein of the reticuloendotheliosis virus (REV),the CA gene of REV DBYR1102 strain was cloned into prokaryotic expression vector pET-30a (+),and recombinant plasmid pET30a-CA was constructed.Mter double enzyme digestion and sequencing,the pET30a-CA was transformed into the Escherichia coli BL21 (DE3).The transformants were induced by isopropylthio-β-D-galactoside (IPTG),and SDS-PAGE was used to identify the expression of the CA protein.The His-tagged fusion protein REV-CA was ultrasonically treated and then purified through Ni-chelating affinity chromatography and sugar concentration.The purified CA was used to immune into New Zealand rabbit for preparing anti-CA polyclonal serum.The results showed that the recombinant plasmids pET30a-CA was successfully constructed,and the CA protein was expressed in soluble form.Polyclonal antibody was prepared and purified from immunized New Zealand white rabbit serum with the titer of 1 ∶ 25 600.This study laid the foundation for the developmem of the diagnostic methods and subunit vaccine for REV.关键词
禽网状内皮组织增生症病毒/CA/原核表达/抗体Key words
avian reticuloendotheliosis virus/CA/prokaryotic expression/purification/antibody分类
农业科技引用本文复制引用
姜莉莉,王立成,陈强,樊兆斌..禽网状内皮组织增生症病毒CA蛋白的表达纯化及多克隆抗体制备[J].畜牧与兽医,2017,49(9):67-70,4.基金项目
国家自然科学基金项目(31402170) （31402170）
辽宁省自然科学基金项目(2014022045) （2014022045）
菏泽学院博士基金(XY16BS09) （XY16BS09）
