华南地区粉尘螨主要变应原Derf2的cDNA克隆及序列分析

目的克隆并分析华南地区粉尘螨主要变应原Derf2的cDNA片段。方法广州地区收集、鉴定、培养的粉尘螨,提取总RNA,RT-PCR扩增其Derf2片段,连接入T载体,测序和分析。结果Derf2片段长度为558bp,与GenBank公布的Derf2(D10448)的核酸序列比较,在62个核苷酸处插入了87个核苷酸,插入点前后的核酸序列没有差异,按原读码框进行读码显示插入了29个氨基酸,插入点前后的氨基酸序列没有改变。结论获得华南地区粉尘螨的Derf2cDNA片段,和已公布的序列相比,有较大差异。

cDNA Cloning and Sequence Analysis of Major Allergen of Dermatophagoides farinae Derf2 in South China

Objective To clone and analyze the cDNA of major allergen Der f 2 of Dermatophagoides farinae in south China. Methods cDNA of Der f 2 was cloned by RT-PCR, screened and their sequences were analyzed. Results cDNAof Der f 2 was cloned. The sequence of the cloned Der f 2 was different with that published (D10448) in GenBank, with 87 additional nucleotides inserted into the 62th nucleotide of the original one. According to the original ORF, the deduced amino acids that were located prior or after the inserted 29 amino acid sequence showed no changes. Conclusion The cDNA of Der f 2 was cloned from Dermatophagoides farinae and its sequence showed significant difference with that reported in the GenBank.