粉尘螨变应原第6组分基因克隆及序列多态性分析  

Cloning of the gene encoding the group 6 allergen of Dermatophagoides farinae dust mites and analysis of its sequence polymorphisms

作　　者：夏伟 易忠权 赵盼雯 崔玉宝 XIA Wei;YI Zhong-quan;ZHAO Pan-wen;CUI Yu-bao(Jiangsu Vocational College of Medicine,Yancheng 224000,Jiangsu,China;Department of Central Laboratory,The Third People＇s Hospital of Yancheng City;Department of Clinical Laboratory,Wuxi People＇s Hospital Affiliated to Nanjing Medical University)

机构地区：[1]江苏医药职业学院,江苏盐城222400 [2]盐城市第三人民医院中心实验室 [3]无锡市人民医院检验科

出　　处：《中国病原生物学杂志》2018年第9期994-997,共4页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.NSFC31572319)

摘　　要：目的获得粉尘螨变应原第6组分(Der f 6)的编码基因并了解其序列多态性。方法根据GenBank(AF125187)公布的Der f 6的CDS区序列设计引物,使用PrimeSTAR?HS DNA进行PCR扩增,将目的基因片段与pMD19-T simple连接,进行序列测定和多态性分析。结果获得粉尘螨Der f 6编码基因,大小为839bp。对5个Der f 6克隆质粒测序并与参考序列GenBank No.AF 125187和GenBank No.EU085359比对,出现突变的cDNA序列合计36处,cDNA克隆中序列存在27处突变至少达到3个;推导的氨基酸序列有15处与上述27个位点一致。结论获得粉尘螨Der f 6编码基因并证明其序列具有多态性,为相关研究奠定了基础。Objectives To obtain the gene encoding the group 6 allergen of Dermatophagoides farinae（Der f 6）and ascertain its sequence polymorphisms. Methods The total RNA of D.farinae was extracted,and the CDS sequence was amplified with PCR using PrimeSTAR HS DNA in accordance with the sequences in GenBank.The target gene fragment was ligated into a pMD19-T plasmid,sequenced,and its sequence polymorphisms were analyzed. Results The gene coding for Der f 6 was 839 base pairs（bp）in length.Its sequences were compared to reference sequences（GenBank No.AF 125187 and GenBank No.EU0853595）.DNA sequencing of Der f 6 indicated that 27 of 36 mismatched nucleotides were conserved,and amino acids at 15 sites were consistent with the 27 sites mentioned. Conclusion The gene coding for Der f 6 was obtained.This gene displayed polymorphism,providing a basis for further research.

关 键 词：粉尘螨变应原第6组分 序列多态性 基因克隆 变态反应性疾病 

分 类 号：R384.4[医药卫生—医学寄生虫学]