Prokaryotic expression of LIF gene of giant panda and preparation and characterization of monoclonal antibodies
In order to obtain the giant panda LIF monoclonal antibody,the coding region 23-203 gene fragment of giant panda LIF gene was synthesized by adding enzyme cutting sites and ligated into pET30a prokaryotic expression vector,the recombinant plasmid was transformed into BL21 competent cells,the protein was induced by IPTG and purified as an immunogen to immunize BALB/c mice.The hybridoma cells were prepared by conventional method,and the positive hybridoma cell lines were screened and identi-fied by indirect ELISA.The results showed that the LIF(23-202)-pET30a recombinant plasmid was success-fully constructed,recombinant LIF protein immunogen with high purity was obtained.A hybridoma cell line secreting giant panda LIF monoclonal antibody was obtained,and the antibody subtype was IgG2b.The results of Western-blot showed that LIF-2 monoclonal antibody could specifically detect LIF pro-tein in endometrial stromal cells,testicular Sertoli cells,skin cells,umbilical cord mesenchymal stem cells and bone marrow mesenchymal stem cells of giant panda.These results laid an important foun-dation for elucidating the localization and tissue expression information of LIF and exploring the role of LIF in the implantation of giant panda embryos.
国家科技期刊平台
NSTL
万方数据
