水产学报2017，Vol.41Issue(4)：498-505,8.DOI:10.11964/jfc.20160610454
紫贻贝LC3B蛋白的原核表达及抗血清制备
Recombinant expression and polyclonal antibody preparation of Mytilus galloprovincialis LC3B
摘要
Abstract
Autophagy is a cellular process for degradation of damaged proteins and organelles via forming autophagosome.Microtubule-associated protein 1 light chain 3 B (LC3B) is a marker protein for autophagy detection.However,in many lower animals,autophagy detection is limited for a lack of available specific LC3B antibody.In the present study,the coding sequence of LC3B ofMytilus galloprovincialis was cloned and inserted into a pET-32a prokaryotic expression vector.To obtain a high-level and soluble expression of recombinant MgLC3B-His protein,the IPTG concentration and induction time were investigated.Then,the purification conditions of this recombinant protein were also studied.Our results showed that MgLC3B was inserted in the pET-32a prokaryotic expression vector successfully.The recombinant MgLC3B-His protein was induced at 20 ℃,150 r/min with IPTG concentration of 0.6 mmol/L after 10 h culture.A single-band recombinant protein was obtained after affinity purification.After immune injection,and the rabbit polyclonal antibody was also obtained,with a titer of 25 600.Therefore,our results will be helpful for the investigation of autophagy in mussels and similar species.关键词
紫贻贝/LC3B/重组表达/细胞自噬/多克隆抗体Key words
Mytilus galloprovincialis/LC3B/recombinant expression/autophagy/polyclonal antibodies分类
生物科学引用本文复制引用
余振兴,朱倩,姚翠鸾..紫贻贝LC3B蛋白的原核表达及抗血清制备[J].水产学报,2017,41(4):498-505,8.基金项目
国家自然科学基金(41276178,41076076)National Natural Science Foundation of China (41276178,41076076) （41276178,41076076）
