Expression of 3-hydroxyacyl-CoA dehydrogenase in E.coli and preparation of its antiserum
- VernacularTitle:3-羟酰基辅酶A脱氢酶在大肠杆菌中的表达及抗血清的制备
- Author:
Changyuan WANG
;
Chuang XU
;
Binglei SHEN
- Publication Type:Journal Article
- Keywords:
HCDH;
Prokaryotic expression;
Antiserum
- From:
Chinese Journal of Immunology
1985;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To clone and express the HCDH gene fragment in E.coli,and then to prepare its antiserum.Methods:The HCDH gene fragment was amplified by PCR,and ligated to the expression vector pMAL-P2X after DNA sequencing analysis.Recombinant plasmid was transformed into the E.coli.Then its expression was induced by IPTG and analyzed by SDS-PAGE.The otter rabbit was immunized with maltose resin affinity purified antigen.Antiserum valence was detected using ELISA and AGP test.The specificity was detected using Western blot.Results:The objective products of 786 bp were obtained,then the prokaryotic expression vector of HCDH gene was established and the corresponding antiserum was prepared successfully.Conclusion:It lays solid foundation for the further researches on detecting the changes of HCDH protein on the expression level or the expression regulation of HCDH gene.
