Cloning and sequencing of allergens of dermatophagoides (house dust mite)
Isolated DNA encoding allergens of Dermatophagoides (house dust mites) particularly of the species Dermatophagoides farinae and Dermatophagolides pteronyssinus, which are protein allergens or peptides which include at least one epitope of the protein allergen. In particular, DNA encoding two major D. farinae allergens, Der f I and Der f II and DNA encoding a D. pteronyssinus allergen, Der p I. In addition, the proteins or peptides encoded by the isolated DNA, their use as diagnostic and therapeutic reagents and methods of diagnosing and treating sensitivity to house dust mite allergens.
1 . (canceled)
2 . A method of producing an isolated protein of Dermatophagoides pteronyssinus comprising the steps of:
a) culturing a host cell transformed with a nucleic acid encoding a protein allergen of Dermatophagoides pteronyssinus, Der p I, comprising the amino acid sequence shown in FIG. 7 , in an appropriate medium to produce a mixture of cells and medium containing said protein allergen; and
b) purifying said mixture to produce isolated Der p I protein allergen.
3 . The method of claim 2 wherein the nucleic acid comprises a nucleotide sequence selected from the group consisting of the nucleotide sequence shown in FIG. 7 , the coding region of the nucleotide sequence shown in FIG. 7 , and a nucleotide sequence which hybridizes under high or low stringency conditions to a complementary strand of the nucleotide sequence shown in FIG. 7 .
4 . The method of claim 3 , wherein the high stringency conditions comprise hybridization at Tm-20 followed by at least one post-hybridization wash at Tm-12.
5 . The method of claim 2 , wherein the nucleic acid comprises the cDNA insert of phage clone λgt11 p1(13T), ATCC Deposit No. 69338.
6 . A method of producing an isolated protein of Dermatophagoides pteronyssinus comprising the steps of:
a) culturing a host cell transformed with a nucleic acid encoding a protein allergen of Dermatophagoides pteronyssinus, Der p I, and selected from the group consisting of the nucleotide sequence shown in FIG. 7 , the coding region of the nucleotide sequence shown in FIG. 7 , and a nucleotide sequence which hybridizes under high or low stringency conditions to a complementary strand of the nucleotide sequence shown in FIG. 7 , in an appropriate medium to produce a mixture of cells and medium containing said protein allergen; and
b) purifying said mixture to produce isolated Der p I protein allergen.
7 . A method of producing an isolated antigenic peptide of Dermatophagoides pteronyssinus comprising the steps of:
a) culturing a host cell transformed with a nucleic acid encoding an antigenic peptide of Dermatophagoides pteronyssinus, Der p I, comprising a portion of the amino acid sequence shown in FIG. 7 , in an appropriate medium to produce a mixture of cells and medium containing said peptide; and
b) purifying said mixture to produce isolated Der p I peptide.
8 . The method of claim 7 wherein the nucleic acid comprises a portion of a nucleotide sequence selected from the group consisting of the nucleotide sequence shown in FIG. 7 , the coding region of the nucleotide sequence shown in FIG. 7 , and a nucleotide sequence which hybridizes under high or low stringency conditions to a complementary strand of the nucleotide sequence shown in FIG. 7 .
9 . The method of claim 7 , wherein the high stringency conditions comprise hybridization at Tm-20 followed by at least one post-hybridization wash at Tm-12.
10 . The method of claim 7 , wherein the nucleic acid comprises a portion of the cDNA insert of phage clone λgt11 p1(13T), ATCC Deposit No. 69338.
11 . A method of producing an isolated protein of Dermatophagoides pteronyssinus comprising the steps of:
a) culturing a host cell transformed with a nucleic acid encoding an antigenic peptide of Dermatophagoides pteronyssinus, Der p I, and selected from the group consisting of a portion of the nucleotide sequence shown in FIG. 7 , a portion of the coding region of the nucleotide sequence shown in FIG. 7 , and a portion of the a nucleotide sequence which hybridizes under high or low stringency conditions to a complementary strand of the nucleotide sequence shown in FIG. 7 , in an appropriate medium to produce a mixture of cells and medium containing said protein allergen; and
b) purifying said mixture to produce isolated Der p I protein allergen.