Eriksson T L, Whitley P, Johansson E, van Hage-Hamsten M, Gafvelin G
Department of Clinical Immunology, Karolinska Institute and Hospital, Stockholm, Sweden.
Int Arch Allergy Immunol. 1999 Aug;119(4):275-81. doi: 10.1159/000024204.
Dust mites are a major cause of allergic disease worldwide. The dust mite Acarus siro is an inducer of occupational allergy among farmers, but sensitisation has also been found in non-farming populations.
A degenerate primer was designed to the N-terminal amino acid sequence of a 15-kD IgE-binding protein in A. siro extract. The cDNA sequence was obtained by using reverse transcriptase polymerase chain reaction, standard cloning and sequencing techniques. The protein was expressed in Escherichia coli with a 6-histidine tag at its C-terminus. Immunoblotting of the recombinant protein and whole extract was performed using patient sera.
15 and 17-kD allergens were identified in a fraction of A. siro extract. The cDNA of the 15-kD allergen was isolated, cloned and sequenced and the allergen was expressed as a recombinant protein. The calculated molecular weight of the cDNA-encoded protein is 14.2 kD. The predicted amino acid sequence has one potential N-glycosylation site at position 4-6 and a cytosolic fatty acid-binding protein signature at position 5-22. The protein has 64% sequence identity with Blo t 13, an allergen from the dust mite Blomia tropicalis, as well as homology with several other fatty acid-binding proteins (FABPs) from different organisms. The allergen was named Aca s 13 and was recognised strongly by 3 of 13 (23%) of the subjects investigated. The amino acid sequence of the 17-kD protein was partly determined and it also showed high sequence homology with Blo t 13 and FABPs.
尘螨是全球过敏性疾病的主要病因。粉尘螨是农民职业性过敏的诱发因素，但在非农业人群中也发现了致敏情况。
根据粉尘螨提取物中一种15-kD IgE结合蛋白的N端氨基酸序列设计了简并引物。通过逆转录聚合酶链反应、标准克隆和测序技术获得cDNA序列。该蛋白在大肠杆菌中表达，其C端带有6个组氨酸标签。使用患者血清对重组蛋白和全提取物进行免疫印迹分析。
在粉尘螨提取物的一部分中鉴定出15-kD和17-kD过敏原。分离、克隆并测序了15-kD过敏原的cDNA，并将该过敏原表达为重组蛋白。cDNA编码蛋白的计算分子量为14.2 kD。预测的氨基酸序列在第4-6位有一个潜在的N-糖基化位点，在第5-22位有一个胞质脂肪酸结合蛋白特征序列。该蛋白与热带无爪螨的过敏原Blo t 13有64%的序列同一性，并且与来自不同生物体的其他几种脂肪酸结合蛋白（FABP）具有同源性。该过敏原被命名为Aca s 13，在所研究的13名受试者中有3名（23%）能强烈识别它。部分确定了17-kD蛋白的氨基酸序列，它也与Blo t 13和FABP显示出高度的序列同源性。
