Background Information

Question 1

Reasons for biochemical manipulation of genes by using enzymes

Answer 1

A).- Facilitate the study of gene expression and physiological
regulation
B).- Identify gene product or bring about the overexpression of the
gene product
C).- Study protein structure/function relationships
D).- Identify interactions between cellular components with
particular nucleic acid sequences or protein domains

Question 2

What is a vector? What are the properties of a vector?

Answer 2

a replicon designed to carry passenger DNA of interest

Properties:
A).- capacity to function in various host organisms
B).- contains selection markers
C).- bring about a high-level expression of gene of interest

Question 3

What is a replicon?

Answer 3

a piece of DNA that is able to function like a chromosome in vivo

Question 4

What is a plasmid? What features does it contain?

Answer 4

Plasmid is a vector, a small circular double-stranded extrachromosomal DNA

features:
-replication origin (ori)
-antibiotic resistance gene
-multiple cloning site (MCS)

Question 5

What is cloning?

Answer 5

introduction of recombinant plasmid into bacterial cell where it
will be replicated (amplified)

Question 6

Steps in a recombinant DNA study (GMO)

Answer 6

1) amplify gene of interest with PCR
2) insert gene into a vector
3) introduce the recombinant DNA into living cells to propagate it

Question 7

Composition of the human genome [How many bp, percentage of exons and introns?]

Answer 7

-2.9 billion bp in 23 pairs of chromosomes
-Only 5% are exons → code for proteins
-95% are introns and non-coding sequences with not known or no function

Question 8

What are non-coding sequences? [How are they replicated, how many times are they replicated, what is an alternative name for them? How much of the genome do they represent?]

Answer 8

-self-replicated
-repeated hundred thousand times
-called selfish or parasitic, since no function except their own reproduction
-Accounts for 20% of the genome

Question 9

Gel electrophoresis [What does it do?]

Answer 9

Used to monitor DNA cleavage, at alkaline pH, DNA and RNA are
negatively charged

Question 12

Lambda Phage [phage vs other vectors, how many bp of phage can be replaced]

Answer 12

▪ They enter bacteria more easily
than plasmids
▪ 48 kb DNA fragment of λ Phage
is not essential for productive
infection and can be replaced
by foreign DNA

Question 13

Infection modes of Lambda Phage [Lytic and Lysogenic pathway]

Answer 13

Lytic pathway (breaks cell)
-viral functions are fully expressed, viral DNA and proteins are
produced and packaged into viral particles, leading to the lysis of the host cell
and the appearance of virions

Lysogenic pathway (no break)
-the phage DNA becomes inserted into the host-cell
genome and can be replicated together with the host-cell DNA for many
generations, remaining inactive