Bio exam 4

• By restriction enzymes
• Bacterial enzymes that cut DNA in a sequence specific manner

A small double stranded circular DNA molecule that can replicate independently of the main bacterial chromosome.

Isolate it from other genes of the organism and amplify it to obtain many identical copies

Restriction enzymes are enzymes that are bacterial in origin that can be used to cut DNA at specific sequences so it can be manipulated in the laboratory

Plasmids are small double stranded DNA Molecules found in bacteria that cane replicated independently of the main chromosome (and can therefore accumulate in large number in cell)

• Cloning involves cutting the DNA from the organism of inters with restriction enzyme
• Attach them to plasmids that have been opened at one location by a restriction enzyme.
• The joining is accomplished by DNA Ligase

• After the DNA is cut by DNA Ligase and attached to plasmids the resulting plasmids a re then reintroduced into bacteria.
• The bacteria are spread on agate plates and allowed to grow to form colonies overnight.
• Each colony consists of millions of genetical identical bacteria all derived from single bacterial cell that started growing at that location the night before.
• All bacteria present in a colony contain the same cloned gene that is now isolated from all other genes in the cell.

• A faster way to isolate and amplify a gene is with PCR.
• The target DNA is heated to separate the strands.
• Primers are then added along with DNA polymerase and four nucleotides.
• The reaction is cooled to allow the primers to bind and the DNA polymerase then copies of each of the strands
• Now you have 2 molecules insead of the 1 that you started with.
• Cycle is then repeated
• With each cycle, the amount of DNA doubles 1 to 2 to 4 to 8 to 16……

Methods for determining the order go the nucleotide bases

Polymerase chain reaction

• Helps us identify new genes and their functions
• Revels stretches of DNA involved in the regulation of gene expression (Such as promoters and enhancers

• The number of tandem repeated sequences differs at a number of chromosomal locations in each individual
• For this reason, when DNA is is cut with restriction enzymes it generates unique size fragments for each individual.

• Package the DNA so it fits in the Nucleus
• Keep the genes in an accessible state
• Repress gene expression

• Proteins that bind to DNA and regulate gene expression
• Responsible for turning on and off genes (Opening certain his tomes)

DNA sequences that stimulate gene expression via TF binding

Where RNA polymerase binds on DNA to initiate transcription

Transcription factor

A small piece of glass that contain many different genes 100,000 spots with different genes

• Comercial available
• Converts mRNA to cRNA

• Removing the interns from a strip of DNA
• Making mRNA

Blymphocytes

Antibodies bind to foreign molecules

• Segment 1: many variable region segments
• Segment 2: Many J regions
• Segment 3: Many D regions
• Segment 4: 1 Constant region
• These various segments of DNA are shuffled around to make different combinations during the development of B lymphocytes

Single stranded DNA complementary to the ends of the gene