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目的:原核表达人Oncoprotein18(Op18)蛋白,为 制备抗Op18的mAb作准备.方法:从人皮肤组织中用RT PCR扩增Op18cDNA的全长编码序列,克隆入表达载体pR SETA中,构建Op18的高表达工程菌,并以IPTG诱导表达目 的蛋白,通过亲和层析法纯化表达的Op18融合蛋白.结果: 酶切及测序鉴定证明,获得含有目的基因片段的重组质粒,表 达的Op18融合蛋白以可溶性的形式存在.结论:所获Op18 融合蛋白以可溶性的形式存在,为制备其mAb及进一步研究 Op18在创伤愈合及瘢痕形成中的作用打下了基础.
OBJECTIVE: To prokaryotic express human Oncoprotein18 (Op18) protein in preparation of anti-Op18 mAb.Methods: The full-length coding sequence of Op18 cDNA was amplified by RT PCR from human skin tissue and cloned into the expression vector pR SETA to construct Op18 The expression of the target protein was induced by IPTG, and the expressed Op18 fusion protein was purified by affinity chromatography.Results: The recombinant plasmid containing the gene fragment was obtained by restriction analysis and sequencing analysis, and the expressed Op18 fusion protein Soluble form.Conclusion: The obtained Op18 fusion protein exists in soluble form, which lays a foundation for preparing its mAb and further studying the role of Op18 in wound healing and scarring.