Genetic Recombination

Bacterial Genome Asexual Reproduction Conjugation & Transformation

Bacterial Genome
Circular, double stranded Nucleoid region
where DNA is tightly packed Similar to nucleus in eukaryote but not surrounded by a membrane

Genome contains one chromosomal DNA and many plasmids

Plasmids
Small, circular, self-replicating pieces of DNA (separate from the bacterial chromosome) Contain a small number of genes Can incorporate themselves into the bacterial chromosome Episome: genetic elements that can exist either as a plasmid or as part of the bacterial chromosome

Advantage of Plasmids
Plasmids are not required for bacterial cells to survive under normal conditions Under stress, genes on plasmids can confer advantages (e.g. resistance) Plasmids increase genetic variation and thus the likelihood of survival in bacteria

Important Plasmids
R (resistance) plasmid
genes that make bacteria resistant to antibiotics Also carries genes encoding the sex pili

F (fertility) plasmid facilitates genetic recombination

Genes for production of sex pili Fig 18.15a (male = F+, female = F-) female become male if receive F plasmid from male

Asexual Bacterial Reproduction

Binary fission Asexual form of reproduction rapid Bacterial colony = identical clone

Origins of Mutation
Although new mutations are individually rare (low probability of spontaneous mutation) bacterial proliferation (growth rate) is high Thus, new mutations can have a significant impact on genetic diversity when reproduction rate is high Contrast with slow reproductive organisms (e.g. humans) where heritable variation is not due to new mutations but due to sexual recombination of existing genetic information

Mathematical Mutation
E.coli reproduction
~ 2 x 1010 new cells per day

Spontaneous mutation rate

~ 10-7 mutations per division (1 in 10 million)

E. coli mutation rate

2 x 1010 divisions per day x 10-7 mutations per division 2000 bacteria with mutations per day

Thought Question
Can spontaneous mutations alone along with a high reproduction rate account for the diversity seen in bacteria? Experiments: Lederberg and Tatum

Lederberg & Tatum Experiment

(1946)

http://biology-forums.com/gallery/33_27_06_11_10_48_52.jpeg

Hypothesis
If spontaneous mutations was the sole cause of diversity, then after many reproductive cycles:
bacterial mutants that normally can not survive on minimal media should be able to should see living cells in minimal media

Lederberg & Tatum Experiment

(1946)

http://biology-forums.com/gallery/33_27_06_11_10_48_52.jpeg

Example in textbook: Fig 18.12

E. coli need both arg and trp to survive Mutants that cant make arg or trp will die unless provided in the medium
Mutant makes arg not trp Mutant makes trp not arg

Grow on minimal medium

(solution of glucose and salts without arg and trp)

X
No colonies

X
No colonies

Thought Question
What evidence is there that something else helps increase the genetic diversity in bacteria? Experiments:
Lederberg and Tatum Recall: Griffith

Lederberg & Tatum Experiment

(1946)

http://biology-forums.com/gallery/33_27_06_11_10_48_52.jpeg

Lederberg & Tatum Experiment

(1946)

http://biology-forums.com/gallery/33_27_06_11_10_48_52.jpeg

Lederberg & Tatum Experiment

(1946)

http://biology-forums.com/gallery/33_27_06_11_10_48_52.jpeg

Lederberg & Tatum Experiment

(1946)

http://biology-forums.com/gallery/33_27_06_11_10_48_52.jpeg

Example in textbook: Fig 18.12

E. coli need both arg and trp to survive Mutants that cant make arg or trp will die unless provided in the medium Add both mutants together into same minimal medium
None should live but some did!!!
Mutant makes arg not trp Mixture of mutants Mutant makes trp not arg

Grow on minimal medium

(solution of glucose and salts without arg and trp)

No colonies

Colonies

No colonies

Recall: Griffiths Experiment

Recall: Griffiths Experiment

Recombinant

Evidence of Genetic Recombination in Bacteria

Spontaneous mutation alone could not explain why the mixture of cells could survive in minimal medium Cells that survived must have acquired genes from the other mutant strain Even though new mutations are a major source of genetic variation in bacteria, genetic recombination adds more diversity

Mechanism of Genetic Recombination

Eukaryote:
Sexual reproduction / fertilization Crossing over / meiosis

Prokaryotes:
Conjugation Transformation Transduction

Sexual Bacterial Reproduction

Conjugation: direct transfer of genetic material between two joined bacterial cells One way DNA transfer:
male donor female recipient

Mechanism
Donor extends sex pili to recipient Sex pilus retracts, pulling cells together DNA transferred through cytoplasmic bridge
http://static.open.salon.com/files/bacterial_conjugation21228227521.jpg

Conjugation

http://www.pc.maricopa.edu/Biology/rcotter/BIO%20205/LessonBuilders/Chapter%209%20LB/Ch9b9.html

Transformation
Alteration of a bacterial cells genotype by the uptake of naked foreign DNA When two different strains of bacteria are mixed together, eventually they will show characteristics of both strains

Transformation

Conditions for Transformation

Some bacteria have surface proteins that recognize and transport DNA from closely related species Some bacteria only transform when placed in a specific type of environment
E.g. E.coli in high calcium concentration

HW Question
Describe the major similarity between conjugation and transformation. Explain the major difference between what is required for conjugation to occur compared to transformation.

Comparing Mechanism of Bacterial Genetic Recombination

Mechanism Requirement Physical contact via sex pili; F plasmid Free DNA in the environment does not require a cell or cell is dead Killed by bacteriophage State of donor State of recipient

Conjugation

Living

Living

Transformation

Living

Transduction

Bacteriophage

Living