Mice OCILrP2 Cloning prokaryotic expression and antiserum preparation.doc

 PAGE \* MERGEFORMAT 7 Mice OCILrP2 Cloning prokaryotic expression and antiserum preparation Author: Yu Jing-Peng Li, Zhao Yong Jin Di Raoen [Keywords:] mice OCILrP2;, prokaryotic expression;, anti-OCILrP2 antiserum [Abstract] Objective: In the prokaryotic expression system inhibit osteoclasts OCILrP2 a member of the lectin family genes, and produce anti-rat anti-OCILrP2 serum. Methods: RTPCR was cloned from mouse spleen cells, some of OCILrP2 cDNA, recombinant expression vector pET28aOCILrP2, and transformed into E. coli BL21 (DE3), with IPTG induced expression HisOCILrP2 fusion protein. To express the product of the immune SD rats were prepared OCILrP2 anti-rat anti-serum. Was determined using Western blot potency and specificity. Results: RTPCR amplified a 235 bp of the OCILrP2 gene, whose cDNA sequence in GenBank sequence of the same login. To construct the recombinant plasmid was transformed into E. coli pET28aOCILrP2 high expression of target protein. To express the fusion protein of the immune HisOCILrP2 access to anti-rat anti-mouse OCILrP2 serum. Western blot analysis showed that the anti-serum can be expressed with the prokaryotic and eukaryotic HisOCILrP2 fusion protein expression OCILrP2Ig fusion protein-specific binding; anti-serum titer of 1:2000. Conclusion: The expression in prokaryotic cells in mice OCILrP2 protein OCILrP2 prepared anti-rat anti-mouse serum, in order to further study the biological functions of OCILrP2 basis. [Keywords:] mice OCILrP2; prokaryotic expression; anti OCILrP2 antiserum Osteoclast inhibitory lectin-related protein 2 (osteoclast inhibitory lectinrelated protein 2, OCILrP2) is osteoclast inhibitory lectin (OCIL) a member of the family belongs to type Ⅱ transmembrane glycoprotein, with a smaller intracellular area, the extracellular domain containing a C-type lectin domain. Mouse OCILrP2 gene is located on chromosome 6 of the NK gene complex in vivo. This complex, there are many C-type lectin-related genes, incl