CAJ | 학술논문

从深圳地区采集粉尘螨纯培养，提取总RNA，根据香港中文大学合成的尘螨基因序列并设计的引物，RT-PCR扩增出Der f4基因，克隆到pUC57载体后测序和进行生物信息学分析.将该目的基因克隆到pET-28a表达载体上得到重组质粒pET-28a-Der f4.用生物软件分析尘螨变应原Der f4序列. RT-PCR获得目的基因Der f4 cDNA全长为1593 bp.推测编码蛋白由526个氨基酸组成，生物信息学分析表明，Der f4具有多种磷酸化位点，含有信号肽，为疏水性蛋白，包含淀粉酶抑制剂功能结构域.获得Der f4基因全长，其编码的细胞外疏水性蛋白可能具有淀粉酶抑制剂活性.
종심수지구채집분진만순배양，제취총RNA，근거향항중문대학합성적진만기인서렬병설계적인물，RT-PCR확증출Der f4기인，극륭도pUC57재체후측서화진행생물신식학분석.장해목적기인극륭도pET-28a표체재체상득도중조질립pET-28a-Der f4.용생물연건분석진만변응원Der f4서렬. RT-PCR획득목적기인Der f4 cDNA전장위1593 bp.추측편마단백유526개안기산조성，생물신식학분석표명，Der f4구유다충린산화위점，함유신호태，위소수성단백，포함정분매억제제공능결구역.획득Der f4기인전장，기편마적세포외소수성단백가능구유정분매억제제활성.
RNA was extracted from Shenzhen area and the full-length of Der f4 gene was amplified by RT-PCR with the primers designed according to previous sequence published in Hong Kong chinese university. The target gene was cloned into pUC57 plasmid and sequenced Der f4 gene was recombinated into pET-28 a expression vector fol-lowing sequencing identification. And then Der f4 protein was characterized by bioinformatics software. The opening reading frame( ORF)of Der f4 was obtained by RT-PCR and was 1 593 bp in full length. The protein encoded by this sequence was deduced to be consisted with 526 amino acids. Bioinformatics analysis of Der f4 which proved to be hydrophobic proteins associated with amylase inhibitors function structure domain contained signal peptide and multiple phosphorylation site. The full-length cDNA for Der f4 obtained,which should en-code a hydrophobic extra-cellular protein associated with inhibition of amylase.