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Exodus 15:26

Exodus 15:26

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Exodus 15:26

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  1. Exodus 15:26 26 … If thou wilt diligently hearken to the voce of the LORD thy God, and wilt do that which is right in his sight, and wilt give ear to his commandments, and keep all his statutes, I will put none of these diseases upon thee, which I have brought upon the Egyptians: for I am the LORD that healeth thee.

  2. Phage Strategies Timothy G. Standish, Ph. D.

  3. Phage Reproduction:The Lytic Cycle Destruction of the bacteria’s DNA Infection Replication of the viral genome Production of viral parts Packaging Lysis

  4. Phage Reproduction:The Lysogenic Cycle Circularization of phage DNA Infection Many generations of bacteria Temperate phage Exit of phage On to the lytic cycle Integration of phage DNA into the bacterial genome

  5. The l Genome PRE PR/OR PRM Replication PL/OL tRI nutR nutL tL cro CI N CII PR O CIII P Q tR3 Regulation Recombination g Lysis S cos R b A a W xis B C int Nu3 Head Genes att D E FI J FII I K L Z M U H V G T Tail Genes

  6. Order of Gene Expression Immediate Early Replication cro CI N CII O CIII P Q Regulation Recombination g Lysis Delayed Early S cos R b A a W xis B C int Nu3 Late Head Genes att D E FI J FII I K L Z M U H V G T Tail Genes

  7. The l Regulatory Region tR1 tL nutL nutR N cI cro cIII cII Terminator left N utilization left N utilization right Terminator right Positive regulator Anti-terminator Repressor Anti-repressor Positive regulator “c” = clear referring to the clear plaques resulting from functional c genes

  8. The l Regulatory Region PRM PRE PL/OL OR/PR tR1 tL nutL nutR N cI cro cIII cII Promoter Repressor Establish-ment Promoter/ Operator left Promoter Repressor Maintainence Promoter/ Operator right Positive regulator Anti-terminator Repressor Anti-repressor Positive regulator “c” = clear referring to the clear plaques resulting from functional c genes

  9. The l Regulatory Region:Infection PRM PRE PL/OL OR/PR tR1 tL nutL nutR N cI cro STOP STOP cIII cII Transcription Transcription Transcription Transcription Host RNA Polymerase recognizes PL and PR and begins transcription of the N and cro proteins

  10. The l Regulatory Region:Infection - Establishment of Lysogeny PRM PRE PL/OL OR/PR tR1 tL nutL nutR N cI cro cIII cII pN Host RNA Polymerase recognizes PL and PR and begins transcription of the N and cro proteins As they are the very first genes to be expressed, N and cro are called “immediate early” genes. cro

  11. The l Regulatory Region:Infection - Establishment of Lysogeny PRM PRE PL/OL OR/PR tR1 tL nutL nutR N cI cro cIII cII pN pN binds to nut sites allowing transcription through tR1 and tL cro

  12. The l Regulatory Region:Infection - Establishment of Lysogeny PRm PRE PL/OL OR/PR tR1 tL nutL nutR N cI cro pN pN cIII cII Transcription Transcription pN binds to nut sites allowing transcription through tR1 and tL cro

  13. The l Regulatory Region:Infection - Establishment of Lysogeny PRm PRE PL/OL OR/PR tR1 tL nutL nutR N cI cro pN pN cIII cII cII cII cIII cIII The cII and cIII proteins are produced. In vivo a protein called HflA rapidly breaks down cII unless it is combined with cIII. cII up regulates transcription from PRE. cro

  14. The l Regulatory Region:Infection - Establishment of Lysogeny PRm PRE PL/OL OR/PR tR1 tL nutL nutR N cI cro pN pN cIII cII Transcription cII cIII Transcription from PRE makes mRNA complimentary to the cro mRNA thus inhibiting cro translation cro

  15. The l Regulatory Region:Infection - Establishment of Lysogeny PRm PRE PL/OL OR/PR tR1 tL nutL nutR N cI cro pN pN cIII cII Transcription cII cIII The transcript from PRE provides a 3’ untranslated region with an efficient ribosome binding site before the start of the cI gene, thus cI repressor is efficiently produced.

  16. The l Regulatory Region:Infection - Establishment of Lysogeny PRm PRE PL/OL OR/PR tR1 tL nutL nutR N cI cro pN cI cI pN cIII cII cII cIII cI The repressor gene product of cI binds to OR and OL preventing transcription from PR and PL.

  17. The l Regulatory Region:Infection - Establishment of Lysogeny PRM PRE PL/OL OR/PR tR1 tL nutL nutR N cI cro cIII cII Transcription The repressor up-regulates expression from PRM maintaining repressor production and preventing expression of any other proteins and establishing lysogeny. cI cI

  18. The l Repressor Protein Repressor Repressor dimer C C C N N N • Repressor protein has two functional domains: • An amino acid (residue) 1-92 N-terminal DNA binding domain which binds to OR and OL • A C-terminal protein binding domain essential for dimerization running from amino acids 132-236 • A 40 residue connector runs between the two domains Repressor only binds DNA efficiently when it is a dimer

  19. The l Regulatory Region:Infection - Establishment of The Lytic Cycle PRM PRE PL/OL OR/PR tR1 tL nutL nutR N cI cro cIII cII pN Host RNA Polymerase recognizes PL and PR and begins transcription of the N and cro proteins As they are the very first genes to be expressed, N and cro are called “immediate early” genes. cro

  20. The l Regulatory Region:Infection - Establishment of The Lytic Cycle OR/PR PRM Host RNA Polymerase recognizes PL and PR and begins transcription of the N and cro proteins cI cro

  21. The l Regulatory Region:Infection - Establishment of The Lytic Cycle OR/PR PRM OR3 OR2 OR1 Host RNA Polymerase recognizes PL and PR and begins transcription of the N and cro proteins 5’CAT’ACGTTAAATCTATCACCGCAAGGGATAAATATCTAACACCGTGCGTGTTGACTATTTTACCTCTGGCGGTGATAATGGTTGC’ATG3’ 3’ATG’TGCAATTTAGATAGTGGCGTTCCCTATTTATAGATTGTGGCACGCACAACTGATAAAATGGAGACCGCCACTATTACCAACG’TAC5’ cI cro

  22. C C C The l Regulatory Region:Infection - Establishment of The Lytic Cycle N N N OR3 OR2 OR1 5’CAT’ACGTTAAATCTATCACCGCAAGGGATAAATATCTAACACCGTGCGTGTTGACTATTTTACCTCTGGCGGTGATAATGGTTGC’ATG3’ 3’ATG’TGCAATTTAGATAGTGGCGTTCCCTATTTATAGATTGTGGCACGCACAACTGATAAAATGGAGACCGCCACTATTACCAACG’TAC5’ cI cro The N terminus of the repressor binds to OR1 and 2 with high affinity The N terminus of the repressor binds to OR1 and 2 with high affinity The N terminus of the repressor binds to OR1 and 2 with high affinity

  23. The l Regulatory Region:Infection - Establishment of The Lytic Cycle RNA Polymerase RNA Polymerase OR3 OR2 OR1 Host RNA Polymerase recognizes PL and PR and begins transcription of the N and cro proteins 5’CAT’ACGTTAAATCTATCACCGCAAGGGATAAATATCTAACACCGTGCGTGTTGACTATTTTACCTCTGGCGGTGATAATGGTTGC’ATG3’ 3’ATG’TGCAATTTAGATAGTGGCGTTCCCTATTTATAGATTGTGGCACGCACAACTGATAAAATGGAGACCGCCACTATTACCAACG’TAC5’ cI cro

  24. The l Regulatory Region:Infection - Establishment of The Lytic Cycle OR3 CTATCACCGCAAGGGATAA OR2 CTAACACCGTGCGTGTTGA OR1 TTACCTCTGGCGGTGATAA OL3 TAACCATCTGCGGTGATAA OL2 TTATCTCTGGCGGTGTTGA OL1 ATACCACTGGCGGTGATAC OR3 CTATCACCGCAAGGGATAA OR2 CTAACACCGTGCGTGTTGA OR1 TTACCTCTGGCGGTGATAA OL3 TAACCATCTGCGGTGATAA OL2 TTATCTCTGGCGGTGTTGA OL1 ATACCACTGGCGGTGATAC CON -TAYCWCYGGCGGTGWTR- Right and left operator sites share a common consensus sequence.

  25. The End

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