Polymerase Chain Reaction ( PCR )

This is a method to produce very large numbers of copies of specific DNA sequences without cloning. Therefore PCR can amplify specific sequences or add sequences such as  endonuclease  recognition  sequences as primers to cloned DNA.

PCR consists of 5 main components.

-Target DNA

-Single stranded Oligonucleotides (primers)

-d NTPs (dATP, dCTP, dTTP, dGTP)

-Taq DNA polymerase

-Termocycler

There are three main steps in PCR.

Step1:  Denaturation. The mixture of excess primer and DNA fragment is heated to about 95° C causing the double strands of target DNA to be denatured into single strands.

Step 2: Annealing of Primers. The temperature is dropped down between about 35⁰- 65°C

As the temperature decreases the single strands of DNA reassociate into double strands. Large excess of primer allows two primers anneal or bind to their complementary sequences on the target DNA leaving the rest of the fragment single-stranded.

Step 3: Primer Extension. The temperature is raised to 70⁰- 75°C Taq  polymerase is added. Taq polymerase extends the primer into a complementary copy of the entire single-stranded fragment, in the 5’-3’ direction.  As both the DNA strands are replicated, two copies of the original fragment are gained.

This process is repeated many times. At each time, the number of DNA copies doubles. This is continued until enough copies are gained for the analysis. 

Differences between physisorption and chemisorption

Different kinds of bacteriophages (phages)

M13

Filamentous.

About 870nm in length and 6nm in width.

Consists of single stranded DND (ssDNA).

Three kinds of capsomeres build the capside.

Infects E. coli by adsorbing to the cell and entering through F pilli. Therefore only infect F⁺ and HFr cells. Also male specific.

Does not kill the host. Particles are released by budding, therefore when the particles are released, the host cell is alive.

An efficient vector in gene cloning as it can hold longer pieces of foreign DNA.

T phage

Structure is composed of icosahedral head, double stranded DNA and a tail.

Infects E.coli

 Main types of T phages are T2, T4 and T12.

In the infection linear DNA of the phage is released to the host cell and becomes circular by replicating that later produces a long DNA chain known as ‘Concatamen’. This coils into the phage’s head by headfull mechanism while packaging.

Infection kills the host cell as the new particles are released outside by bursting the host cell.

Lambda (λ)

Composed of head and tail.

Head is consisting of double stranded linear DNA.

At both 5’ ends of the DNA strand, 12 complementary base pair, single stranded segments are present. These two ends are known as “cos ends”.

Because of the cos ends, phage chromosome circularizes before replication. Concatamen is produced during the replication and during the packaging, Terminase enzyme cuts off the cos ends.

Host cell is E.coli.

MS2

Contains the smallest known genome.

Super coiled single stranded DNA.

Infect only through sex pilli. Therefore male specific.

Infect E.coli.

Phi×174(ΦX174)

Contains a single stranded circular DNA.

After adsorption, synthesizes the complementary strand and becomes double stranded.

Use as a positive control in DNA sequencing.

G4

Structurally similar to ΦX174 phage.

Can infect susceptible E.coli cells. 

Serotonin the neurotransmitter

Serotonin i.e. 5-hydroxytryptamine (5-HT) is a monoamine neurotransmitter that is said to be helping to relay signals from one area of the brain to another. However its primary functions are found in gastrointestinal tract as 90% of total serotonin is located in the enterochromaffin cells in the gut. 

Serotonin is made via a unique biochemical conversion process that begins with Tryptophan. Tryptophan is a building block to proteins. In the synthesis of serotonin, Tryptophan hydroxylase the enzyme combines with tryptophan to form 5-hydroxytryptophan metabolite that later converts to Serotonin.

Via wikipedia.org

As mentioned earlier serotonin helps to distribute messages across the brain. Brain cells related to mood, appetite, sleep, memory, learning, temperature regulation, sexual desire and some social behaviour are influenced either directly or indirectly by serotonin due to the widespread distribution in the brain. 

It can also affect the functioning of the cardiovascular system, muscles, and various elements in the endocrine system.

When it comes to mental health, it is widely believed that a serotonin deficiency plays a role in depression but there is no way to measure its levels in the living brain. Therefore, there have not been any studies proving that brain levels of this or any neurotransmitter are in short supply when depression or any mental illness develops. People who suffer from depression shows lower serotonin levels in blood levels but still it is not revealed that whether the blood levels reflect the brain's level of serotonin.

Antidepressant medications such as SSRIs (selective serotonin reuptake inhibitors) and SNRIs (serotonin and norepinephrine reuptake inhibitors) that work on serotonin levels are believed to lower the symptoms of depression, but their exact function is not fully understood.

Recent studies show that when the Mycobacterium vaccae, which occurs naturally in soil and is often breathed in when spending much time nature, is injected into mice, it stimulates neuron growth and causes serotonin levels in blood to increase. So that the bacteria could have antidepressant benefits but it is not yet revealed whether it has an effect on human.