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目的 克隆、表达可溶性肝抗原（ＳＬＡ）及细胞色素Ｐ４５０ ２Ｄ６（ＣＹＰ ２Ｄ６）。方法 采用 ＲＴ—ＰＣＲ技术从人肝组织ｐｏｌｙ（Ａ）＋ＲＮＡ中扩增ＳＬＡ及 ＣＹＰ ２Ｄ６ ｃＤＮＡ，经ＢａｍＨ Ⅰ、Ｈｉｎｄ Ⅲ双酶切定向插入载体ＰＱＥ－３０并在大肠杆菌Ｍ１５中表达。对表达载体ＰＱＥ—３０／ＳＬＡ、ＰＱＥ—３０／ＣＹＰ ２Ｄ６中的目的基因进行序列分析，表达产物用 ＳＤＳ—ＰＡＧＥ、免疫印迹方法鉴定。结果 表达产物经 ＳＤＳ－ＰＡＣＥ和免疫印迹分析后在分子量 ４．７ × １０４和５．０ × １０４处各有一条明显的蛋白带，并分别能特异性与抗ＳＬＡ、ＣＹＰ２Ｄ６阳性血清反应。结论 表达ＳＬＡ、ＣＹＰ２Ｄ６为自身免疫性肝炎的诊断及其发病机制研究提供物质基础。
Objective To clone and express soluble liver antigen (SLA) and cytochrome P450 2D6 (CYP 2D6). Methods The cDNA of SLA and CYP 2D6 was amplified from human liver poly (A) + RNA by RT-PCR and inserted into vector PQE-30 by restriction endonuclease BamH Ⅰ and Hind Ⅲ to express in E. coli M15. The target genes in the expression vectors PQE-30 / SLA and PQE-30 / CYP 2D6 were sequenced and the expressed products were identified by SDS-PAGE and Western blotting. Results After SDS-PAGE and Western blot analysis, the expressed products had a distinct protein band at molecular weights of 4.7 × 104 and 5.0 × 104, respectively, and could specifically react with anti-SLA and CYP2D6-positive serum respectively. Conclusion The expression of SLA and CYP2D6 provide the material basis for the diagnosis and pathogenesis of autoimmune hepatitis.