中文摘要：

目的获得粉尘螨变应原第16组分（Der f 16）编码基因并构建其原核表达体系。方法提取粉尘螨总RNA,根据GenBank（AF465625）设计并合成引物,经RT-PCR合成Der f 16全长基因,克隆至pCold TF DNA载体,热转化至E.coli JM109;将测序正确的pCold TF-Der f 16质粒转化BL21（DE3）,IPTG诱导表达,并用SDS-PAGE验证产物。结果 RT-PCR扩增获得目的基因Der f 16,全长1 443bp,与参考序列同源性99.72%。将构建的pCold TF-Derf 16质粒转化宿主菌E.coli BL21后进行诱导,SDS-PAGE电泳显示其全细胞、上清及沉淀物均有蛋白表达,且上清表达量高于沉淀物。生物信息学分析显示该重组蛋白由480个氨基酸组成,分子质量单位为55.1315ku,二级结构由（-螺旋（35.21%）、延伸主链（20.83%）和无规卷曲（43.96%）组成。该变应原含有4个凝溶胶蛋白位点。结论粉尘螨变应原第16组分原核表达获得成功,为进一步规模生产该变应原并应用于临床诊治奠定了基础。

英文摘要：

Objectives To determine the gene coding for the group 16 allergens of Dermatophagoides farinae（Der f 16） and construct a prokaryotic expression system.Methods Total RNA was isolated from D.farinae and used as template for RT-PCR amplification of the gene fragments coding for Der f 16.These fragments were then cloned into a pCold TF DNA vector and transformed into E.coli JM109.After correct sequencing,the plasmid pCold TF-Der f 16 was transformed into E.coli BL21（DE3）,its expression was induced with IPTG,and it was identified with SDS-PAGE.Results The gene coding for Der f 16 was obtained by RT-PCR and was 1 443 bp in full length.It was 99.72% identical to the reference sequence.The constructed plasmid pCold TF-Der f 16 was then transformed into E.coli BL21 and its expression was induced with IPTG.It was then identified with SDS-PAGE.The target protein was expressed in whole cells,supernatant,and precipitate containing pCold TF-Der f 16.Bioinformatics indicated that the recombinant allergen consisted of 480 amino acid residues and had a molecular weight of 55.1315 ku.Its secondary structure consisted of an alpha helix（35.21%）,extended strand（20.83%）,and random coil（43.96%）.The Der f 16 had four gelsolin domains.Conclusions Der f 16 was successfully expressed in E.coli,providing a basis for large-scale production of the recombinant allergen for clinical diagnosis and treatment of allergic disorders.