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Lecture3 - MCB 110L Lecture 3 Qing Zhong 306 Barker Hall...

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MCB 110L Lecture 3 1/26/2011 Qing Zhong 306 Barker Hall
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An open reading frame starts with an atg (Met) in most species and ends with a stop codon (taa, tag or tga). Open reading frame mRNA---cDNA
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Define Full-length cDNA Peptide hits-protein sequence cDNA sequence Search all the cDNA database Hypothetic full-length Open reading frame-start Human/mouse genome -Longest reading frame -stop codon before start codon
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E.coli Expression construct - PET24b Vector Type: Bacterial Stable/Transient: Transient Constitutive/Inducible: Constitutive Expression Level: High Backbone size: 3509 Sequencing Primer: T7 Tag: T7 (Nterm), His (Cterm) Mammalian Selection: Kanamycin Comments:Same as pET21abcd(+) but kanR; a,b,c,d vary by MCS;
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Fusion protein-delete the stop codon Cin8 Promoter 3’ untranslated region Start codon Stop codon Cin8 Promoter 3’ untranslated region Start codon Stop codon Start codon Stop codon GFP GFP
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Kinesin subfamily that contains Cin8p
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Antibody made in rabbits that binds heavy chain of human neuronal kinesin
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Closely related, not identical
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