please help OK So lets go back to CandyP the delicious prote

please help!

OK. So, let\'s go back to CandyP, the delicious protein we talked about in class that tastes like a Snickers bar. You painstakingly derived the transcribed sequence for CandyP and now need to get E. coli to make that protein. What do you need to do to the transcribed sequence before inserting it into a plasmid to make sure that the E. coli will make this protein? Why? Compare and contrast eukaryotic DNA replication bubbles and transcription bubbles. The Sectumsempra toxin binds to dicer and renders it completely non-functional. What effects would this particular toxin have on the rate of translation? Explain. (2-3 sentences) Draw a eukaryotic gene with two exons and include its pre-mRNA and mRNA products. Label all important features. Now that you know about eukaryotic transcripts, how would a prokaryote differ in this process?

Solution

1. After the transcription process is done I need to do the tailoring for the sequence made proper to insert into a plasmid. I need to look into the marker gene that would let me know after inserting the whole plasmid into an E. coli. I have to check for the 3\' and 5\' tails to be fit for knitting it with a plasmid, here I have to use \'Restriction Digestion Endonuclease\' enzyme. This enzyme is very necessary to make the right insertion into a plasmid. So the recognition sequence into an expression vector would help replicate the exact gene.

2. While a cell division occurs both daughter cells needs to get the DNA from mother cell so replication carries out on the other hand DNA decodes to form an RNA through the process of transcription gets us the transcription bubble.

Replication Bubble

Transcription Bubble

Purpose is to take the entire genome to the next progeny.

Here a RNA molecule would be transcribed.

Leading and lagging strand both are used as templates for the replication process.

The strand of DNA in a bubble opens up and one stand helps making the RNA.

One (DNA) Polymerase helps synthesizing.

One (RNA) polymerase helps in synthesizing.

There is a specific origin where the bubble starts in a synthesis process.

Here also the origin is the initiation part making the transcription process done.

Contrasts: Replication Bubble:

i) It gets opened by the helicase,

ii) DNA polymerase helps in replicating it,

iii) Double stranded DNAs are formed by this mechanism.

Transcription Bubble:

i) RNA polymerase helps in the whole process,

ii) RNA polymerase binds to the exposed DNA,

iii) Single stranded RNAs are formed by this mechanism.

3. Dicer molecule helps in RNA silencing complex, it binds to the RNA and makes it single stranded (micro RNA) that would go and bind to a mRNA finally leading to RNA silencing that is translation to stop. If Sectumsempra toxin affects the dicer molecule in that case the translation would not be hindered and it will go on, a negative effect on cell control mechanisms.

4. Prokaryotic transcripts do not possess \'Introns\',

They do not contain any of the 3\' caps or any of the \'Poly A\' tail,

RNA processing part is quiet simpler, there is no modification like splicing and capping unlike eukaryotes,

There is no precursor RNA like eukaryotes.

Replication Bubble	Transcription Bubble
Purpose is to take the entire genome to the next progeny.	Here a RNA molecule would be transcribed.
Leading and lagging strand both are used as templates for the replication process.	The strand of DNA in a bubble opens up and one stand helps making the RNA.
One (DNA) Polymerase helps synthesizing.	One (RNA) polymerase helps in synthesizing.
There is a specific origin where the bubble starts in a synthesis process.	Here also the origin is the initiation part making the transcription process done.