粉尘螨过敏原β-己糖胺酶的表达、纯化和生物信息学分析  

Expression,Purification and Bioinformatics Analysis ofβ-hexosaminidase of Dermatophagoides farinae

作　　者：莫丽华[1] 刘玉琳[2] 杨利桃[3] 范小琴[3] 刘志刚[1] 杨平常[1] 

机构地区：[1]深圳大学过敏反应与免疫学研究所,深圳518060 [2]南昌大学医学院免疫学教研室,南昌330006 [3]深圳大学耳鼻咽喉科研究所,深圳518020

出　　处：《中国寄生虫学与寄生虫病杂志》2015年第2期159-160,F0003,共3页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金(No.31328014;31400786);深圳市南山区研发项目(No.KC2012JSYB0003A)~~

摘　　要：合成粉尘螨β-己糖胺酶基因并连接至p ET-28a载体,用异丙基-β-D-硫代半乳糖苷(IPTG)诱导表达重组β-己糖胺酶蛋白。通过生物信息学软件分析粉尘螨β-己糖胺酶蛋白基本特性。表达菌经诱导后,高效表达出β-己糖胺酶蛋白,SDS-PAGE结果显示表达产物相对分子质量(Mr)约为55 000。粉尘螨β-己糖胺酶基因序列全长1 410 bp,编码469个氨基酸,位于胞外,无信号肽,属亲水性蛋白,其二级结构由14.71%的片层,30.70%的螺旋和54.58%的环组成。The DNA fragment encoding β-hexosaminidase was synthesized， and cloned into pET-28a vector. The constructed plasmid pMD18-T-β-hexosaminidase was transformed into E. coli Top10 and followed by expression of the protein induced by IPTG. SDS-PAGE result showed that the relative molecular mass of the recombinant protein was about Mr 55 000. The full length of β-hexosaminidase gene was 1 410 bp. Bioinformatics analysis revealed that β-hexosaminidase was composed with 469 amino acid residues with a calculated molecular weight of Mr 55 000， and its secondary structure was composed of strand（14.71%）， helix（30.70%）， and loop（54.58%）. β-hexosaminidase was a hydrophilic protein without signal peptide， and located in the extracellular space.

关 键 词：粉尘螨 &beta -己糖胺酶 原核表达 

分 类 号：R384.42[医药卫生—医学寄生虫学]