Cloning and Subcloning of cDNA Coding for Group Ⅱ Allergen of Dermatophagoides Farinae
Abstract：Objective: To clone, sequence and subclone the cDNA coding for group Ⅱ allergen of Dermatophagoides farinae( Der f 2 ). Methods: The cDNA gene fragment of Der f 2 was amplified by RT-PCR. After being purified, the gene fragment was cloned into a vector pMD-18T. The recombinant plasmid pMD-18T-Der f 2 was transformed into E. coli JM109. Positive clones were screened and identified by PCR and digested with restriction endonuclease, and the sequence of inserted Der f 2 eDNA was also analyzed. Then Der f 2 was subcloned into the vector of pET-32a( + ). Results: The Der f 2 eDNA was specifically amplified from RNA by RTPCR. The recombinant plasmid pMD-18T-Der f 2 and pET-32a( + ) -Der f 2 was constructed and digested by Sac I and Not I , and the size of gene fragment was 455bp and in accordance with the expected one. Conclusion: The pET-32a( + )-Der f 2 subclone was constructed successfully.
Keywords：Dermatophagoides farinaehouse dust mitecDNAsequence analysisantigen
Publication Date：2004-08-31
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 239-243 )