cDNA Cloning and Expression of Blo t 11, the Blomia tropicalis Allergen Homologous to Paramyosin
cDNA Cloning and Expression of Blo t 11, the Blomia tropicalis Allergen Homologous to Paramyosin
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Blo t 11（与副肌球蛋白同源的热带布洛米亚过敏原）的 cDNA 克隆和表达
DOI：
10.1159/000049525
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发表时间：
2001
影响因子：
2.8
通讯作者：
K. Chua
中科院分区：
文献类型：
--
作者：
J. D. Ramos;N. Cheong;B. Lee;K. Chua
热带布氏螨是世界上许多地区的重要螨种，也是热带国家最主要的螨种。对该物种致敏的普遍性可能被低估了，因为商业提取物基本上不可用。B的鉴定和表征。tropicalis过敏原是了解该物种在过敏性致敏中的作用的重要一步，并可为诊断和治疗程序提供适当的试剂。本文报道了一个编码A B的cDNA基因的分离、序列分析、表达及变应原性研究。热带过敏原与副肌球蛋白同源，副肌球蛋白是一种高分子量过敏原，以前在粉尘螨中发现。通过cDNA文库筛选、cDNA末端5′快速扩增和长距离PCR，获得了Blo t 11全长cDNA基因。用CLUSTAL W、CGC和BLAST程序包的组合进行序列分析。cDNA基因在大肠杆菌中表达为GST融合蛋白，并通过使用谷胱甘肽琼脂糖柱的亲和层析进行纯化。用人IgE斑点印迹法检测rBlo t 11的致敏性。Blo t 11基因全长3,111 bp，开放阅读框为2,625 bp，编码875个氨基酸，与多种无脊椎动物副肌球蛋白具有很高的同源性。人IgE斑点杂交免疫分析表明，rBlot 11对52%（33/63）的哮喘患者血清呈阳性反应。Blo t 11是Der f 11的同系物，表现出潜在的重要过敏活性。
Blomia tropicalis is an important mite species in many parts of the world and the most predominant mite species in tropical countries. The prevalence of sensitization to this species has probably been underestimated because commercial extracts are largely unavailable. Identification and characterization of B. tropicalis allergens is an important step toward understanding the role of this species in allergic sensitization and could provide appropriate reagents for diagnostic and therapeutic procedures. This paper describes the isolation, sequence analysis, expression and allergenicity of a cDNA gene coding for a B. tropicalis allergen with homology to paramyosin, a high-molecular-weight allergen previously identified in Dermatophagoides farinae. The full-length Blo t 11 cDNA gene was isolated by cDNA library screening, 5′-rapid amplification of cDNA ends and long-distance PCR. Sequence analysis was performed with a combination of CLUSTAL W, CGC and BLAST program packages. The cDNA gene was expressed as a GST fusion protein in Escherichia coli and purified by affinity chromatography using the glutathione Sepharose column. Allergenicity of the rBlo t 11 was tested by human IgE dot blot immunoassay. Blo t 11 is a 3,111-bp cDNA gene with a 2,625-bp open reading frame coding for an 875-amino acid protein, exhibiting significant homology with different invertebrate paramyosins. The human IgE dot blot immunoassay showed that the rBlo t 11 reacted positively to 52% (33/63) of sera from asthmatic patients. Blo t 11 is the homolog of Der f 11 exhibiting potentially important allergenic activity.
通过 cDNA 克隆鉴定热带布洛米亚过敏原 Blo t 5。
影响因子：
2.8
作者：
Arruda,LK;Fernandez-Caldas,E;Naspitz,CK;Montealegre,F;Vailes,LD;Chapman,MD
通讯作者：
Chapman,MD
编码热带布洛米亚过敏原的互补 DNA 的核苷酸序列分析。
DOI：
10.1016/s0091-6749(96)80009-1
发表时间：
1996
期刊：
The Journal of allergy and clinical immunology
影响因子：
--
作者：
Puerta,L;Caraballo,L;Fernández-Caldas,E;Avjioglu,A;Marsh,DG;Lockey,RF;Dao,ML
通讯作者：
Dao,ML
秀丽隐杆线虫副肌球蛋白 N 末端区域的磷酸化。
影响因子：
5.6
作者：
Schriefer,LA;Waterson,RH
通讯作者：
Waterson,RH