【摘要】 通过原核表达Mig-14蛋白,制备相应多克隆抗体,用于后续Mig-14分子功能研究,为探寻Mig-14拮抗PB分子机制奠定基础.经密码子偏爱性分析显示mig-14基因含有8%的稀有密码子,综合考虑密码子偏好性、GC含量、限制性酶切位点等影响因素对密码子进行优化,重新合成mig-14基因,PCR获得周质空间部分,克隆至表达载体pET22b(+),并转入大肠埃希菌JM109中表达,KCl染色后切胶纯化的蛋白作为抗原免疫家兔,制备多克隆抗体.Mig-14蛋白多克隆抗体的制备为后续免疫共沉淀研究Mig-14的作用蛋白奠定了基础.更多还原

【Abstract】 In order to explore the molecular function of Mig-14,we try to express Mig-14and prepare the anti-Mig-14polyclonal antibody.Codon preference analysis showed mig-14gene contains 8%of rare codons.In this study,mig-14was synthesized by considering the codon bias,GC cotent,restriction site et al.mig-14in the periplasm which acquired by PCR using synthesized mig-14as template was cloned into pET22b(+)and expressed in E.coli JM109.The recombinant protein Mig-14-p(Mig-14in the periplasm)was purified by cutting the gel slices with KCl stain and was used as antigen to prepare polyclonal antibody.This anti-Mig-14-p polyclonal antibody can be used to research the role of mig-14in Salmonella enterica serovar Typhi.更多还原