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目的构建一个能在原核高效表达人白细胞介素－１３（ＨＩＬ－１３）的系统。方法采用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）获得ＨＩＬ－１３的ｃＤＮＡ，将其克隆至原核表达载体ｐＧＥＸ－２Ｔ后进行诱导表达。结果合成了特异编码ＨＩＬ－１３的ｃＤＮＡ，构建了原核表达质粒ｐＧＥＸ－２ＴＨＩＬ－１３，对其进行序列分析表明，表达质粒中的插入片段为ＨＩＬ－１３基因；诱导表达出含有ＨＩＬ－１３的融合蛋白谷光苷肽－Ｓ－转移酶－ＨＩＬ－１３（ＧＳＴ－ＨＩＬ－１３），其相对分子质量为３９０００。结论（１）从Ｔ淋巴细胞中克隆出ＨＩＬ－１３基因。（２）ＨＩＬ－１３ｃＤＮＡ在原核细胞表达的蛋白占菌体总蛋白的３７％。
Objective To construct a system capable of efficiently expressing human interleukin-13 (HIL-13) in prokaryotes. Methods The cDNA of HIL-13 was obtained by reverse transcription-polymerase chain reaction (RT-PCR) and cloned into prokaryotic expression vector pGEX-2T for inducing expression. Results The cDNA encoding HIL-13 was synthesized and the prokaryotic expression plasmid pGEX-2THIL-13 was constructed. The sequence analysis showed that the inserted fragment in the expression plasmid was HIL-13 gene. The fusion protein containing HIL-13 Protein glutathione-S-transferase-HIL-13 (GST-HIL-13), the relative molecular mass of 39000. Conclusion (1) HIL-13 gene was cloned from T lymphocytes. (2) HIL-13 cDNA expressed 37% of the total protein in the prokaryotic cells.