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- Title
栗酒裂殖酵母 SOT22 蛋白全长和N端的原核表达与多克 隆抗体制备.
- Authors
刘金王; 黄 英
- Abstract
To bioinformatically analyse the biological characteristics of Schizosaccharomyces pombe SpTrz2 protein, both the full-length and the N-terminal half of S. pombe sptrz2 gene were amplified by PCR and inserted into pET-28a ( + ) expression vector through Nde I and Xho I restriction enzyme sites. The recombinant plasmids were transformed into Escherichia coli BL21 ( DE3) and the transformants were induced by IPTG. Then recombinant proteins were purified by the way of SDS-PAGE gel slices . Polyclonal antibodies were prepared by immunizing New Zealand white rabbit using the purified recombinant proteins. The specificity of antibodies was tested by Western blotting. The results showed that SpTrz2 was a transmembrane protein with three transmembrane domains and contained multiple B cell antigen epitopes. SDS-PAGE analysis suggested that the full-length and the N-terminal half of SpTrz2 protein ( SpTrz2 and SpTrz2N ) with a relative molecular mass of about 75. 99 ku and 44. 77 ku, respectively, mainly existed in a form of inclusion body. The Western blot analysis showed that polyclonal antibodies of SpTrz2 and SpTrz2N could specifically recognize natural SpTrz2 protein. In this study, recombinant SpTrz2 and SpTrz2N protein were successfully expressed and purified, and their poly clonal antibodies were prepared. All above results could provide experimental evidence for the study of biological function of SpTrz2 protein.
- Publication
Acta Agriculturae Zhejiangensis, 2021, Vol 33, Issue 1, p34
- ISSN
1004-1524
- Publication type
Article
- DOI
10.3969/j.issn.1004-1524.2021.01.05