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  • Title: [Cloning, sequencing and subcloning of cDNA coding for group I allergen of Dermatophagoides farinae].
    Author: Yang QG, Li CP.
    Journal: Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi; 2004 Jun; 22(3):173-5. PubMed ID: 15597716.
    Abstract:
    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. CONCLUSION: The pET-32a(+)-Der f 1 subcloning has been constructed successfully.
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