病毒学英文版经典课件Virology-To-Students.ppt

* * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * Example for Primer Design: AGATACATCTGGAGACTTTCGGAAAGCCTTGCTCCGGCTCCGGATCGGACGATTGCGTCGCATCGACCCTGCGCCCAAGCTGCATCATCGAAATTGCCGTCAACCAAGCTCTGATAGAGTTGGTCAAGACCAATGCGGAGCATATACGCCCGGAGCCGCGGCGATCCTGCAAGCTCCGGATGCCTCCGCTCGAAGTAGCGGCCATTGTCCGTCAGGACATTGTTGGAGCCGAAATCCGCGTGCACGAGGTGCCGGACTTCGGGGCAGTCCTCGGCCCAAAGCATCAGCTCATCGAGAGCCTGCGCGACGGACGCACTGACGGTGTCGTCCATCACAGTTTGCCAGTGATACACATGGGGATCAGCAAGGCCATGGTTGTGTATCTCTAAGAATCTAAGAA Upper A2 BamH I: 5’-AAAAAAGGATCCAGATACATCTGGAGACTTTCGGAAAGCCTTGCT-3’ (Length: 45bp, TM: 75?C, GC: 42.2%) Lower A2 Kpn I: 5’-CCCACCGGTACCTTCTTAGATTCTTAGAGATACACAACCATGGCC-3’ (Length: 45bp, TM: 74?8?C, GC: 48.9%) * The sequences of the primers and linker: Hu Vh back (23bp) 5’-CAGGTGCAGYTRNDGSAGTCDGS-3’ Hu Jh forward (24bp) 5’-TGAGGAGACGGTGACCRKKGTBCC-3’ Hu Vh back Sfi I Nco I (56bp) 5’- GTCCTCGCAACTGCGGCCCAGCCGGCCATGGCCCAGGTGCAGYTRNDGSAGTCDGS-3’ Hu Jh forward Not I (48bp) 5’-GAGTCATTCTCGACTTGCGGCCGCTGAGGAGACGGTGACCRKKGTBCC-3’ Hu Vk back (23bp) 5’-GAMATYSWGMTSACBCAGTCTCC-3’ * Hu Vλ back (23bp) 5’-YMBKYTRTRYTGACKCARBMNBC-3’ Hu Jk forward Not I (48bp) 5’-GAGTCATTCTCGACTTGCGGCCGCACGTTTGATYTCCASYYKKGTCCC-3’ Hu Jλ forward Not I (48bp) 5’-GAGTCATTCTCGACTTGCGGCCGCACCTARRACGGTSASSTKGGTCCC-3’ Linker (45bp) 5’-GGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCG-3’ Reverse Jh for ScFv linker (40bp) 5’-GVACMMYGGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGG-3’ Reverse Vk for ScFv linker (42bp) 5’-GGAGACTGVGTSAKCWSRATKTCCGATCCGCCACCGCCAGAG-3’ Reverse Vλ for ScFv linker (42bp) 5’-GVNKVYTGMGTCARYAYARMVKRCGATCCGCCACCGCCAGAG-3’ * Fc V H L CDR FR A model of IgG molecule * A linker will link HV and LV together A. Recombined ScFv gene insert B. Recombined ScFv antibody Primer VH Linker VL A B * Peptide (12mers) NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN Randomly synthesized peptide gene insert Terminals linked to the MCS of phage Peptide gene i