背景:城市地区最常见的蟑螂类型之一,美洲蟑螂(美洲大鹦鹉),据报道,过敏和哮喘的风险增加。由于缺乏基因组相关信息,在该物种中已鉴定出有限的过敏原组(每1-13)。
方法:为了扩大美洲疟原虫的过敏原分布,基因组,转录组,并应用了蛋白质组学方法。在使用纳米孔组装的高质量基因组的支持下,Illumina,和Hi-C测序技术,根据蛋白质同源性鉴定潜在的过敏原。然后,使用酶联免疫吸附测定,在对美洲疟原虫过敏的泰国患者中测试了选定的过敏原。
结果:美洲疟原虫(3.06Gb)的染色体水平基因组已组装,具有94.6%的BUSCO完整性,其连续性得到了显著提高(N50=151Mb)。全面的过敏原概况已被表征,有七组新的过敏原,包括烯醇化酶(每a14),细胞色素C(每15),科菲林(每a16),α-微管蛋白(每17),亲环蛋白(每18),porin3(每19),和过氧化物酶6(每20),在酶联免疫吸附试验中显示IgE敏感性。使用生物信息学和蛋白质组学方法揭示了一种新的原肌球蛋白同变应原(每7.02)和每5的多种潜在同变应原。此外,美洲疟原虫与密切相关的Blattodea物种的比较分析揭示了交叉反应的可能性。
结论:美洲疟原虫的高质量基因组和蛋白质组有利于在分子水平上研究蟑螂过敏原。在Pera7中鉴定了七个新的变应原组和一个等变应原。
One of the most common cockroach types in urban areas, the American cockroach (Periplaneta americana), has been reported to impose an increased risk of allergies and asthma. Limited groups of allergens (Per a 1-13) have been identified in this species due to the lack of genome-related information.
To expand the allergen profile of P. americana, genomic, transcriptomic, and proteomic approaches were applied. With the support of a high-quality genome assembled using nanopore, Illumina, and Hi-C sequencing techniques, potential allergens were identified based on protein homology. Then, using enzyme-linked immunosorbent assay, selected allergens were tested in Thai patients allergic to P. americana.
A chromosomal-level genome of P. americana (3.06 Gb) has been assembled with 94.6% BUSCO completeness, and its contiguity has been significantly improved (N50 = 151 Mb). A comprehensive allergen profile has been characterized, with seven novel groups of allergens, including enolase (Per a 14), cytochrome C (Per a 15), cofilin (Per a 16), alpha-tubulin (Per a 17), cyclophilin (Per a 18), porin3 (Per a 19), and peroxiredoxin-6 (Per a 20), showing IgE sensitivity in enzyme-linked immunosorbent assay. A new isoallergen of tropomyosin (Per a 7.02) and multiple potential isoallergens of Per a 5 were revealed using bioinformatics and proteomic approaches. Additionally, comparative analysis of P. americana with the closely related Blattodea species revealed the possibility of cross-reaction.
The high-quality genome and proteome of P. americana are beneficial in studying cockroach allergens at the molecular level. Seven novel allergen groups and one isoallergen in Per a 7 were identified.