Microbial Genetics Flashcards
The two parental strands reassociate after acting as templates for new strands, thus restoring the parental double helix
Conservative model of DNA.
The two strands of the parental molecule separate, and each functions as a template for synthesis of a new, complementary strand
Semiconservative model of DNA.
Each strand of both daughter molecules contains a mixture of old and newly synthesized DNA.
Dispersive model of DNA.
Showed that DNA replicates in a semi-conservative way (1958)
The Meselson-Stahl experiment
abundant, basic proteins with a positive charge that bind to DNA.
5 main types: H1, H2A, H2B, H3, H4
~equal in mass to DNA
evolutionarily conserved
Histone-charged proteins
Packing of DNA into chromosomes:
1- Winding of DNA around histone octamer to create a nucleosome structure
2- Nucleosomes connected by strands
of linker DNA like beads on a string.
3- Packaging of nucleosomes into
30-nm chromatin fiber.
4- Formation of looped domains.
Transformation
The genetic alteration of a bacterial cell resulting from the direct uptake and incorporation of exogenous (outsider) genetic material from its surroundings through the cell membrane
Plasmid
Naturally occurring circular dsDNA molecules.
Separated from the main genomic DNA of organism.
Exist in multiple copies (a few to 100’s) in bacterial cell, 1 kb to 200 kb.
Single, circular, double stranded DNA.
Function of plasmid
-Enhancement of adaptation to bacterial environment by transferring genes horizontally (i.e., without normal conjugation).
-Responsible for out break of antibiotic resistant pathogenic bacteria, due to transfer of plasmids containing antibiotic resistance genes between bacterial spp.
Enzyme activity can be used as marker for gene insertion
Disrupted gene = nonfunctional
Intact gene = functional
Media containing XGAL chromagenic substrate used (blue colonies = intact; white colonies = disrupted)
Enzyme beta galactosidase (lac Z)
Operon
A set of adjacent structural genes, plus the adjacent regulatory signals that affect transcription of the structural genes.
Bacterial conjugation
It’s the process by which one bacterium transfers genetic material to another through direct contact. During conjugation, one bacterium serves as the donor of genetic material, and the other serves as the recipient
“F” Plasmid
‘fertility factor’ confers donor characteristics through (sex pilus) to bacterial cells.
Bacteria that have F plasmid are referred to as F+ or Donor.
Those that do not have F plasmid are called F- or recipient.
Tra-region
Genes coding the F-Pilus and DNA transfer process.
OriT
The sequence which marks the starting point of conjugative transfer
Agrobacterium
a genus of bacteria that causes tumors in plants
Ti Plasmid consists of two main regions
T-DNA gene region
Virulence gene region
Encodes for plant growth hormones including auxins and cytokinins, which promotes cell division and induce tumor formation (Crown gall disease)
T-DNA gene region
The bacterial genes in operons VirA-VirE are necessary for DNA transfer to take place
Virulence gene region
-Wound sites on plants attract Agrobacterium through the release of phenolic compounds.
Chemotactic response in Agrobacterium T-DNA transfer
VirA
Transmembrane receptor, which detects the plant aceto-syringone from the wound site
VirG
“Transcription activator, in which it activates the rest of the Vir operons”.
VirC
ssDNA synthesis of the T- DNA
VirD1
Nick the T-DNA at the L and R boarders