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Cloning a Histidine‐rich Loop of the CrMTP1 Protein of Chlamydomonas reinhardtii
Author(s) -
Yang Huidong,
Gingrich David J
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.699.13
Subject(s) - chlamydomonas reinhardtii , chlamydomonas , cloning (programming) , histidine , biology , zinc , biochemistry , gene , zinc finger , transport protein , transmembrane protein , chemistry , microbiology and biotechnology , amino acid , receptor , organic chemistry , mutant , computer science , transcription factor , programming language
Zinc plays several roles in cellular metabolism. To accomplish this, the amount and forms of zinc must be controled within the cell. One important aspect of zinc homeostatis is the transport of zinc ions across cell membranes via transport proteins. This research involves the cloning of a loop of a putative zinc‐transport protein CrMTP1from the organism Chlamydomonas reinhartdii . This histidine‐rich region of the protein, between putative transmembrane helicies 4 and 5, may provide zinc binding sites involved directly in zinc transport or in regulation. PCR has been utilized to amplify the complete gene from a Chlamydomonas cDNA library and subsequently the sequence of the gene corresponding to only the histidine loop region. The PCR amplification is designed to incorporate the DNA into an expression vector in order to produce the protein in E. coli . After purification, putative zinc binding to the purified protein will be studied in order to investigate its possible role in the function of the putative zinc transport protein.