动物医学进展Issue(11)：29-33,34,6.
山羊IFN-γ基因的克隆表达与多克隆抗体制备
Cloning and Expression of Goat IFN-γGene and Preparation of Polyclonal Antibodies
摘要
Abstract
To obtain the expression products of goat IFN-γ,total RNA was extracted from activated goat PBMCs,and cDNA was synthesized and used as template for PCR.The recombinant cloning vector was constructed.The sequence encoding mature peptide region was cloned by PCR and inserted into the ex-pression vector pET-32a,and the recombinant plasmid was transformed into the competent cell BL21 (Co-don Plus),followed by sequencing and analysis.After induced by IPTG,the expression products were and analysized by SDS-PAGE and purified by Ni-NTA.The purified protein was immunize rabbits to prepare polyclonal antibodies.Sequence analysis suggested that sequence encoding the mature peptide was 432 bp. SDS-PAGE indicated that the fusion protein was about 34.9 ku,and a large amount of soluble protein could be expressed when induced with a total concentration of 0.3 mmol/L/L IPTG for 6 h at 30℃.The titter of polyclonal antibodies was 1∶106 detected by indirect ELISA.关键词
山羊/IFN-γ/基因克隆/原核表达/抗体制备Key words
goat/IFN-γ/gene clone/prokaryotic expression/polyclonal antibody分类
农业科技引用本文复制引用
安贝,赵玄多,杨雯昱,侯伟杰,高洋,陈德坤..山羊IFN-γ基因的克隆表达与多克隆抗体制备[J].动物医学进展,2014,(11):29-33,34,6.基金项目
陕西省科技计划项目 （）
