HS BIO 002 - BIOTECHNOLOGY

biotechnology

technology based on biology.

use of theories and concepts in biology science to improve our quality of life.

simply applied biology.

recombinant dna

products of recombination.

genetic engineering

goal is to identify traits that are favorable and insert it in another organism.

can have animals or plants with desirable traits or that they can produce specific protein.

clustered regularly interspaced short palindromic repeats

most common gene editing tool.

bacillus thuringiensis

example of a product of genetic engineering.

type of bacteria that lives in the soil.

producing proteins that are harmful to insects specially several species of moths and butterfly.

target dna

the gene of interest.

donor

the cell organism where the target dna can be found.

restriction enzymes

used to cut dna into fragments.

based from bacteria that are immune to bacteriophages.

naturally 4-8 bases which are palindromic in nature.

may leave sticky ends or blunt ends.

restriction sites

cuts specific segments of the dna.

bacteriophages

virus that infects the bacteria.

dna cloning vectors

carry target gene into host cell, example plasmid and bacteriophage.

carry out dna molecules.

not capable of manifesting in the trait nor produce the protein.

plasmid

is a circular double stranded dna in bacteria.

host cell

bacterial cell that allows the cloning vector to replicate within it.

host should be non-pathogenic, harmless micro-organism which is easy for cultivation.

its common host is escherichia coli.

modifying enzymes

introduces minor bases into dna or rna e.g, dna ligase and taq polymerase.

cloning

a molecular biology technique that makes identical copies of a piece of dna, such as gene.

molecular cloning

involves introducing a specific piece of dna (most often a gene) into a cell where it does not generally belong.

somatic cell nuclear transfer

dolly the sheep, a by product of a reproductive cloning through this primary laboratory process.

reproductive cloning

is a biotechnological process that creates a genetically identical copy of a multicellular organism.

it involves transferring the nucleus from a somatic (body) cell of a donor into an enucleated egg cell, which is then stimulated to develop into an embryo.

dna extraction

extract dna from a cell first, before it starts with cloning.

isolation of a target gene

can be done using (1) cutting the gene from a complete chromosome using restriction enzyme or (2) producing a complementary dna (cdna).

must first be cut intro fragments and the one containin gthe desired gene must be identified.

remember to use the same restriction enzyme.

insertion of a target gene into vector

gene is inserted into vectors such as plasmid and bacteriophages resulting in recombinant dna.

bacterial plasmids that have been isolated from bacterial cells must be mixed with the same restriction enzymes used to cut the dna molecule.

done to produce the same sequence of sticky ends.

introduction of a vector into a host

the plasmids carrying the target gene must be introduced into host cell through transformation, and only lesser amount will contain the recombinant plasmid.

amplification of the target gene by the host cell

bacteria will then be cultured in a medium.

the transformed bacteria will be able to grow and form colonies on the medium.

the bacteria will divide to produce new identical bacterial cells.

each time the bacterial cells divide, the recombinant plasmids will produce multiple copies of the target gene.

polymerase chain reaction

a method used to amplify dna.

resembles the replication but is carried out in a test tube.

thermal cycler

a laboratory instrument that heats and cools samples in repetitive cycles to facilitate dna or rna amplification through the polymerase chain reaction.

denaturation

in which double-stranded dna templates are heated to separate the strand; the reaction temperature is increased to 95 degree celsius, which melts (disrupts the hydrogen bonds between complementary bases) all dsdna into single- stranded dna (ssdna).

annealing

the dna molecule will be exposed at a lower temperature to allow the primers to attach to the single-strand dna.

primer

its role is to let the taq polymerase copy the segment.

taq polymerase

an enzyme derived from a bacteria thermos aquaticus.

mimics the job of dna polymerase but can withstand a high temperature.

extension

temperature will be raised again.

taq polymerase will copy and texted the segment of dna.

after third step, it is considered that 1 cycle is complete and there is 22 amount of dNA in container (usually an eppendorf). so after 1 cycle we can assume that there is 4 copies of dna.