Prokaryotic expression and polyclonal antibody preparation of rat neuroglobin gene
- VernacularTitle:大鼠神经球蛋白的原核表达及多克隆抗体制备
- Author:
Ji ZHU
- Publication Type:Journal Article
- Keywords:
neuroglobin;
prokaryotic expression;
polyclonal antibody;
eukaryotic expression
- From:Journal of Third Military Medical University
1983;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone rat NGB gene and prepare its polyclonal antibody in order to investigate the function of neuroglobin (NGB) Methods The total RNA was extracted from Wistar rat brain and the full length cDNA encoding NGB was obtained by RT-PCR Confirmed by sequencing analysis,it was inserted in the prokaryotic expression vector pET-28a(+) pET-28a(+)-NGB was expressed in Ecoli BL21 after its sequence and reading frame were confirmed by two restriction endonucleases and sequencing The protein of pET-28a(+)-NGB was used to immunize New Zealand rabbits for preparing polyclonal antibody after identification and purification The antibody titer was tested by ELISA method Results The titer of polyclonal antibody was 100 000 Immunoblot analysis showed the polyclonal antibody can recognize fusion protein expressed in 293 cells Conclusion The polyclonal antibody of NGB is prepared and can be used in further study
