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Generation of recombinant proteins was conducted using the methylotrophic yeast Pichia pastoris (P. pastoris) expression system, generally according to Kasuto and Levavi-Sivan [2] and Yom-Din et al. [3], and described in detail in Burow et al. [1]. Synthesis of genes for medaka fshb (Accession Number NM_001309017.1), lhb (Accession Number AB541982.1), fshba, and lhba (gpa; Accession Number NM_001122906) was outsourced to GenScript, New Jersey, U.S.A. For each construct gene expression cassettes were generated with P. pastoris codon optimized DNA sequence. Polyclonal antisera against recombinant mdFshβ and mdLhβ were produced following a procedure according to Aizen et al. [4], which is reported in detail in Burow et al. [1]. |
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Protocol tips |
Generation of recombinant proteins was conducted using the methylotrophic yeast Pichia pastoris (P. pastoris) expression system, generally according to Kasuto and Levavi-Sivan [2] and Yom-Din et al. [3], and described in detail in Burow et al. [1]. Synthesis of genes for medaka fshb (Accession Number NM_001309017.1), lhb (Accession Number AB541982.1), fshba, and lhba (gpa; Accession Number NM_001122906) was outsourced to GenScript, New Jersey, U.S.A. For each construct gene expression cassettes were generated with P. pastoris codon optimized DNA sequence. Polyclonal antisera against recombinant mdFshβ and mdLhβ were produced following a procedure according to Aizen et al. [4], which is reported in detail in Burow et al. [1]. |
Publication protocol
Generation of recombinant proteins was conducted using the methylotrophic yeast Pichia pastoris (P. pastoris) expression system, generally according to Kasuto and Levavi-Sivan [2] and Yom-Din et al. [3], and described in detail in Burow et al. [1]. Synthesis of genes for medaka fshb (Accession Number NM_001309017.1), lhb (Accession Number AB541982.1), fshba, and lhba (gpa; Accession Number NM_001122906) was outsourced to GenScript, New Jersey, U.S.A. For each construct gene expression cassettes were generated with P. pastoris codon optimized DNA sequence. Polyclonal antisera against recombinant mdFshβ and mdLhβ were produced following a procedure according to Aizen et al. [4], which is reported in detail in Burow et al. [1].
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