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1. MICROBIAL GENETICS CONJUGATION (MATING) AND TRANSDUCTION (PHAGE-LIKE TRANSDUCING PARTICLES MOVE DNA). 2. MICROBIAL GENETICS - CONJUGATION. MALE, FERTILITY FACTOR, DONOR FEMALE, ABSENCE OF F FACTOR, RECIPIENT

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Presentation Transcript
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1

MICROBIAL GENETICS

CONJUGATION (MATING)

AND TRANSDUCTION

(PHAGE-LIKE TRANSDUCING PARTICLES MOVE DNA)

slide2

2

MICROBIAL GENETICS - CONJUGATION

MALE, FERTILITY FACTOR, DONOR

FEMALE, ABSENCE OF F FACTOR, RECIPIENT

F+ MALE, SEX PILI, CONJUGATION BRIDGE, ORIGIN OF TRANSFER, NICKING AT ORIT, TRANSFER OF SINGLE STRAND, COMPLEMENTARY STRAND SYNTHESIS IN MALE AND FEMALE

HFR MALE (HIGH FREQUENCY OF RECOMBINATION), INTEGRATION OF TRANSFERRED DNA INTO RECIPIENT CHROMOSOME BY HOMOLOGOUS RECOMBINATION, SELECTION OF RECOMBINANTS

F' MALE, ILLEGITIMATE RECOMBINATION, PARTIAL DIPLOIDS

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3

CONJUGATION - MATING OF MALE AND FEMALES

E. COLI

MALES: FERTILITY FACTOR F PLASMID

~100,000 BASE PAIRS

MALE CHARACTERISTICS

CONTACT WITH FEMALE

DNA DONATION

FEMALES: F -

DNA RECIPIENT

ABSENCE OF F FACTOR

F FACTOR - DOUBLE STRAND DNA

IS2

INSERTION SEQUENCE 1

F

TRANSFER GENES

REPLICATION GENES

oriT ORIGIN OF TRANSFER

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4

F+ MALE

F PILUS

BINARY FISSION

CHROMOSOME AND F FACTOR DUPLICATED

slide5

5

F FACTOR TRANSFER: F+ x F - F+ AND F+

F+

F -

NICK F AT oriT

TRANSFER ONE STRAND

CONJUGATION BRIDGE

NEW SYNTHESIS=

F+

F+

slide6

APPRECIATE !!

TRANSFER GENES OF F FACTOR

slide7

APPRECIATE !

F PILUS SECRETION

slide8

APPRECIATE !

DNA SECRETION APPARATUS

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6

HFR [HIGH FREQUENCY RECOMBINANT]

MALES TRANSFER CHROMOSOME FRAGMENTS

FORMING HFR

F INTEGRATES INTO CHROMOSOME;

F+ BECOMES HFR

oriT

IS2

F+ MALE

PRO+

LAC+

IS2

HOMOLOGOUS

RECOMBINATION

BETWEEN IS2

oriT

IS2

IS2

HFR MALE

PRO+

LAC+

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7

HFR x F- RECOMBINANTS

HFR PRO+ STRS x F- PRO- STRR

RECOMBINANT PRO+ STRR

PROLINE

F-

NEW SYNTHESIS

HFR

CONJUGATION

BRIDGE BREAKS

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8

CONSIDER RECIPIENT CELL ONLY

HOMOLOGOUS

RECOMBINATION -

TWICE LEFT AND

RIGHT OF

MUTATION

LINEAR

FRAGMENT

DEGRADATION

PRO+

STR-R

F-

RECOMBINANT

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9

DEMONSTRATION

GLUCOSE AND

STREPTOMYCIN

~108 HFR

PRO+ STR-S

~108 F-

PRO- STR-R

MIX

~108 HFR

~108 F-

PRO+ STR-R

RECOMBINANTS

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HFR CAN EXCISE F FACTOR PRECISELY

TO REGENERATE F+ MALE

IS2

IS2

HFR

IS2

IS2

HOMOLOGOUS

RECOMBINATION

BETWEEN IS2's

IS2

F+ MALE

IS2

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11

F' MALE : F PLUS CHROMOSOME GENE(S)

PRO+

HFR

LOOPING

STR-S

PRO+

ILLEGITIMATE

RECOMBINATION

DELETION

PRO+

F' MALE

STR-S

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12

F' MATING F' x F- F' + NEW F'

PRO+

PRO+

PRO-

STR-S

STR-R

F' STR-S PRO+

F- PRO- STR-R

PRO+

PRO+

PRO-

STR-S

STR-R

NEW F' MALE

PARTIAL DIPLOID