Respected readers, authors and reviewers, you can add comments to this page on any questions about the contribution, review, editing and publication of this journal. We will give you an answer as soon as possible. Thank you for your support!
CHEN Neng-xing, WEI Xiao-jing, LIU Wei, HOU Lian-sheng. Clone, expression of allantoicase gene from Dictyostelium disc-oideum and preparation of polyclonal antibody against ALC[J]. Journal of East China Normal University (Natural Sciences), 2010, (4): 77-84.
Citation:
CHEN Neng-xing, WEI Xiao-jing, LIU Wei, HOU Lian-sheng. Clone, expression of allantoicase gene from Dictyostelium disc-oideum and preparation of polyclonal antibody against ALC[J]. Journal of East China Normal University (Natural Sciences), 2010, (4): 77-84.
CHEN Neng-xing, WEI Xiao-jing, LIU Wei, HOU Lian-sheng. Clone, expression of allantoicase gene from Dictyostelium disc-oideum and preparation of polyclonal antibody against ALC[J]. Journal of East China Normal University (Natural Sciences), 2010, (4): 77-84.
Citation:
CHEN Neng-xing, WEI Xiao-jing, LIU Wei, HOU Lian-sheng. Clone, expression of allantoicase gene from Dictyostelium disc-oideum and preparation of polyclonal antibody against ALC[J]. Journal of East China Normal University (Natural Sciences), 2010, (4): 77-84.
The coding region of allCis obtained from total mRNA of Dictyostelium discoideumby RT-PCR. The results show that the length of allC is 1 100 bp with sequence analysis and coded protein is 42 kD.Becase it is a differentially expression fragments, indicating that the gene play an important role in multicellular development and allC was cloned into the fusion expression vector pET-32a(+) with 6 His tagged and expressed in E. coli BL21 (DE3) host cells. and then purified by Ni2+ affinity chromatography column and fusion protein purified was used to immune the New Zealand rabbits for preparing polyclonal antibody. The fusion protein was successfully expressed and polyclonal antibody was also successfully obtained. The potency of the antibody was as high as 1∶64 000. The specificity of antibody was proved by Western Blotting analysis of expression product of allC. These data suggest ALC fusion protein has the good antigenicity. The antibody with high titer and specificity was obtained in the satisfaction of Western blot and localization experiment request , which was the foundation to farther study the characters of the allantoicase protein and their function during Dictyostelium discoideumdevelopment.