[Cloning, sequencing and subcloning of cDNA coding for group I allergen of Dermatophagoides farinae].
OBJECTIVE
To clone, sequence and subclone the cDNA coding for group 1 allergen of Dermatophagoides farinae (Der f 1).


METHODS
The cDNA of Der f 1 was amplified by RT-PCR and PCR. After purified, the gene fragment was cloned into a vector pMD-18T. The recombinant plasmid pMD-18T-Der f 1 was transformed into E. coli JM109. Positive clones were screened and identified by PCR and digestion with restriction enzyme. The sequence of inserted Der f 1 gene fragment was also detected. Der f 1 was then subcloned into the vector of pET-32a(+).


RESULTS
The Der f 1 gene fragment of Dermatophagoides farinae was specifically amplified from RNA by RT-PCR and PCR. The recombinant plasmid pMD-18T-Der f 1 and pET-32a(+)-Der f 1 was constructed and digested by Bam H I and Sac I, the size of gene fragment was 646 bp and in accordance with the expected one.


CONCLUSIONS
The pET-32a(+)-Der f 1 subcloning has been constructed successfully.
Yang
Qing-gui
QG
School of Medicine, Anhui University of Science & Technology, Huainan 232001, China.
Li
Chao-pin
CP
D000485:Allergens / Q000235:genetics*; D000818:Animals; D039741:Antigens, Dermatophagoides / Q000235:genetics*; D060829:Arthropod Proteins; D001483:Base Sequence; D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008925:Mites / Q000235:genetics*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics; D017421:Sequence Analysis
D000485:Allergens; D039741:Antigens, Dermatophagoides; D060829:Arthropod Proteins; D018076:DNA, Complementary; D011994:Recombinant Proteins; D003546:Cysteine Endopeptidases; C451159:Dermatophagoides farinae antigen f 1