Sarma P V, Purkayastha S, Madan T, Sarma P U
Department of Biochemical Technology, Sri Venkateswara College, Dhaula Kuan, New Delhi, India.
Immunol Lett. 1999 Dec 1;70(3):151-5. doi: 10.1016/s0165-2478(99)00140-6.
The gene for an 18 kD allergen/cytotoxin of Aspergillus fumigatus was cloned in pUC-19 vector and expressed in Escherichia coli JM109. Digestion of this gene with AluI resulted in four fragments of 216bp, 120bp, 39bp and 21bp. These fragments were cloned in the Sma-I site of pUC-19. The recombinants thus, generated after transformation in E. coli JM109, were screened using monoclonal antibodies raised against the AspfI. The fusion protein containing 120 bp AluI fragment was recognised by the MoAb indicating presence of epitope(s) in the 120 bp region. The study indicates a viable strategy for identification and expression of an immunologically active domain of a major allergen/antigen of A. fumigatus for the first time.
烟曲霉18 kD变应原/细胞毒素的基因被克隆到pUC - 19载体中，并在大肠杆菌JM109中表达。用AluI消化该基因产生了四个片段，分别为216bp、120bp、39bp和21bp。这些片段被克隆到pUC - 19的Sma - I位点。在转化大肠杆菌JM109后产生的重组体，使用针对AspfI产生的单克隆抗体进行筛选。含有120 bp AluI片段的融合蛋白被单克隆抗体识别，表明在120 bp区域存在表位。该研究首次表明了一种可行的策略，用于鉴定和表达烟曲霉主要变应原/抗原的免疫活性结构域。
