XII: Chapter 11- Biotechnology: principles and processes Flashcards
Name the two core techniques that enabled birth of biotechnology.
Genetic engineering and bioprocess engineering
The construction of the first recombinant DNA emerged from the possibility of linking a gene encoding antibiotic resistance with a native plasmid of
Salmonella typhimurium
Which scientists first accomplished the construction of artificial recombinant DNA molecule and when?
Stanley Cohen and Herbert Boyer in 1972
When were the two enzymes responsible for restricting the growth of bacteriophage in Escherichia coli isolated?
1963
What did the two enzymes responsible for restricting the growth of bacteriophage in Escherichia coli isolated do?
One of them added methyl groups to the DNA and the other cut DNA.
Which was the first restriction endonuclease to be isolated/
Hind II
How many restriction nucleases have been identified and from how many strains of bacteria?
900 restriction nucleases from 230 strains of bacteria
Which sequence is responsible for controlling the copy number of the linked DNA?
Origin of replication
How many recognition sites should a (recombinant technology) vector preferably have?
1
In heat shock method of making host competent in genetic engineering, at which temperature are the host cells placed with rDNA after keeping them on ice?
42 celsius
What are primers used in PCR technology?
Small chemically synthesised oligonucleotides that are complementary to the regions of DNA.
In how many cycles of PCR can DNA be amplified a billion times?
30 cycles
What quantities of culture can a bioreactor hold?
100 to 1000L
Detergents are added during purification of DNA to
Break plasma membrane
Which compound is required for the functioning of restriction endonuclease II?
Mg2+
Which palindromic sequence does BamHI recognise?
GGATCC
Which palindromic sequence does Hind III recognise?
AAGCTT
Sal-I restriction endonuclease is extracted from which microbE?
Streptomyces albus
Alu-I restriction endonuclease is extracted from which microbe?
Arthrobacter luteus
What is the function of rop in plasmid vectors?
Codes for proteins required for replication of plasmids
How many cloning sites does pBR322 have?
8
What do the letters stand for in pBR322?
p- plasmid
B- Bolyver
R- Rodrigues
322- experiment number
What is the maximum insert size for plasmid vector?
15kb
WHat is the maximum insert size for lambda phage vector?
23kb
What is the maximum insert size for M-13 phage vector?
25-35 kb
Cosmid vector is obtained from
Cos site of lambda phage
What is the maximum insert size for BAC vector?
200-300 kb
BAC- Bacterial artificial chromosome.
What is the maximum insert size for YAC vector?
1000 kb
YAC- yeast artificial chromosomes
What is the maximum insert size for HAC vector?
> 1000 kb
HAC- human artificial chromosome
Which type of electrophoresis is used for the separation of proteins?
Proteins- SDS PAGE (Sodium dodecyl sulphate polyacryl acryl amide gel electrophoresis
What is used as a dye in SDS PAGE?
Bromophenol blue + toluidine blue
At which end of target DNA do primers attach in PCR?
3’ end
Which vector is made of single stranded DNA?
M-13 phage
