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    • Feature Klfd selectively binds to the CACCC boxes in the promoters of both α-globin and miR-144 genes to activate their transcriptions.
    Application WB
    Species Zebrafish
    Targets Klfd
    Catergory Transcription related
    Title Mir-144 selectively regulates embryonic alpha-hemoglobin synthesis during primitive erythropoiesis.
    Publication Liu TX, Blood, 2009
    Methods A rabbit polyclonal antiserum against zebrafish Klfd was generated using a His-Klfd (N-terminal) fusion protein as an antigen source. A 5' fragment was amplified by polymerase chain reaction (PCR) from the full-length klfdcDNA. The resulting PCR product of 810 bp encoding the amino-terminal 270 amino acids of the Klfd polypeptide was digested with BamH1/Xho1 and subcloned into the BamH1/Xho1 sites of the plasmid pET-28a(+) vector.AHis-Klfd (N-terminal) fusion protein of the expected size (34 kDa) was expressed in Escherichia coli strain DH5 , purified with Ni-NTA agarose (QIAGEN, Valencia, CA), and used to immune a rabbit (AbMART, Shanghai, China).
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