Yuuki T, Okumura Y, Okudaira H
Central Research Laboratories, Asashi Breweries Ltd., Tokyo, Japan.
Int Arch Allergy Immunol. 1997 Jan;112(1):44-8. doi: 10.1159/000237430.
We amplified genomic DNA encoding the major house dust mite allergen, Der f2, from Dermatophagoides farinae by means of the polymerase chain reaction and cloned it into Escherichia coli. The nucleotide sequences of the amplified fragments were determined and compared with those of the cDNA previously reported. Four Der f2 genomic clones were obtained, suggesting that the genomic Der f2 gene had sequence polymorphisms like the cDNA clones. Each of the genomic clones had a single small intron. With respect to the exon sequences, two of the four genomic clones were identical with two cDNA clones, respectively. The others were combinations of the two clones. Genomic Southern blotting suggested that the Der f2 gene is located at one locus in the mite genome and that sequence substitutions were due to polymorphisms among individual mite genes.
我们通过聚合酶链反应从粉尘螨中扩增出编码主要屋尘螨变应原Der f2的基因组DNA,并将其克隆到大肠杆菌中。测定扩增片段的核苷酸序列,并与先前报道的cDNA序列进行比较。获得了四个Der f2基因组克隆,表明基因组Der f2基因具有与cDNA克隆类似的序列多态性。每个基因组克隆都有一个小内含子。关于外显子序列,四个基因组克隆中的两个分别与两个cDNA克隆相同。其他的是这两个克隆的组合。基因组Southern印迹分析表明,Der f2基因位于螨基因组的一个位点,序列替换是由于个体螨基因之间的多态性所致。