粉尘螨Der f14基因克隆与分子特征的分析

从纯培养的粉尘螨中,根据本课题组前期合成的尘螨基因序列,将扩增的Der f14分别克隆到pUC57载体中,经扩增后用SmaI双酶切将目的片段连接到 pET-28a表达载体上得到重组质粒pET28a- Der f14.在线软件ExPaSy,NCBI网站对测序结果进行分析.分析结果表明:获得的目的基因Der f14 cDNA全长为5 013 bp.编码蛋白由1 666个氨基酸组成.粉尘螨Der f14与屋尘螨序列比对同源性达95；,信号肽序列存在1～18个氨基酸,二级结构是由a螺旋(35.83；)﹑延伸链(17.17；)﹑无规则区(47；)组成.目的蛋白是一种卵黄蛋白原.

The Cloning and Bioinformatics Analysis of Der f14 Gene from the Dermatophagoide Farinae

Genes sequence which have been synthesized from the pure culture of dust mites according to the earlier stage of our research group.The Der f14 gene fragment was amplified and cloned into pUC57 vector,and then subcloned into the expression vector pET with restriction enzyme.The recombinant plasmid pET28a-Der f14 was constructed.Then the sequencing result was analyzed by software in ExPaSy,and NCBI web.Results show that the opening reading frame(ORF)of Der f14 was obtained and was 5013 bp in full length.The protein encoded by this sequence was deduced to be consisted with 1 666 amino acids.Similarity of the amino acid sequence of the group 14 allergens were 95； between dermatophagoides farinae and dermatophagoides pteronyssinu.one signal peptide from 1 to 18 position.The secondary structure was composed ofalpha helix(35.83；)extended strand(17.17；)and random coil(47；).The gene of target protein encode a kind of vitellogenin.